accelerating results - affymetrix

USB Product Catalog

PCRGenomics

Custom Manufacturing

responsive

ISO certified

diagnostics

life science research

Accelerating ResultsReagents. Enzymes. Biochemicals.

proven

© 2014 Affymetrix Inc. All rights reserved. FOR RESEARCH USE ONLY.

Affymetrix, Inc. USB® Products26111 Miles RoadCleveland, Ohio 44128 Tel: 888-362-2447 | 216-765-5000Fax: 800-535-0898 | 216-464-5075 [email protected] usb.affymetrix.com

Affymetrix UK Ltd. USB® ProductsVoyager, Mercury Park,Wycombe Lane, Wooburn Green,High Wycombe HP10 0HH, United KingdomTel: +44 (0)1628 55 2600Fax: +44 (0)1628 55 [email protected]

Affymetrix Pte Ltd USB® Products7 Gul Circle, #2M-01 Keppel Logistics Building Singapore 629563 Tel: +65 63957301Fax: +65 63957300 [email protected]

CATALOG ISSUE 1.14

Results.Reagents. Enzymes. Biochemicals.

usb.affymetrix.com

Life Science Research For more than 40 years, USB has been supplying labs worldwide with molecular biology reagents, enzymes, and biochemicals. We offer a wide variety of products that scientists count on to deliver reliable, repeatable results. Our products work the first time so you can confidently advance your research.

Diagnostics We meet the regulatory compliance needs of our diagnostic customers with our ISO 13485 certified site. Our processes are suited for IVD assays and kits, with validated equipment and production practices. We offer rigorous change control capabilities and comprehensive documentation. Top diagnostic companies include our products in their kits and workflows, and in their service labs.

Custom & Bulk Manufacturing From our site in Cleveland, Ohio, we can customize any USB product to your specific need. Our expert staff can assist with bulk sizes, specialized packaging, or custom formulations. Let us lead the manufacturing, labeling, and kitting processes for all of your OEM components.

Product Code Index 252-270

Nucleotides 54-61

PCR Tools 6-53

Molecular Biology Enzymes 62-95

Molecular Biology Products & Kits 96-137

Biochemicals 182-251

Purification 138-155

RNA Analysis 156-167

DNA Sequencing 168-181

Custom, Bulk, and OEM4-5

Product Name Index 271-292

USB Product Catalog

ABOUT USB PRODUCTS

Welcome to our latest catalog. We hope it continues

to be a valued resource.

We’re committed to helping academic institutions,

diagnostic, biotech, and pharmaceutical companies

worldwide excel within the life sciences industry.

Our team is creative, dedicated, and passionate about

working together to achieve success. Our attention

to quality is evident in all that we do, from product

development and operations to our exceptional

technical support and customer service.

In addition to our catalog products, we can customize

our enzymes and biochemicals to fit your specific

requirements. Whether you need a bulk size or a

custom formulation, our expert staff will work within

our rigorous quality system to ensure that your needs

are met.

For the latest information on new products, or to

download protocols, information sheets, or MSDS

documents, please visit us at usb.affymetrix.com.

2 888-362-2447 | 216-765-5000 | usb.affymetrix.com

Ordering Information/ISO Certification

Ordering Information

Orders:Phone: 1-888-362-2447 or 216-765-5000

(8:30 a.m. - 5:30 p.m., E.S.T.)

Fax: 800-535-0898 or 216-464-5075

Website: usb.affymetrix.com

E-mail: [email protected]

Mail: Affymetrix, Inc. USB Products 26111 Miles Road Cleveland, Ohio 44128

In Europe:Phone: +44(0)1628 55 2600

Fax: +44(0)1628 55 2675

Website: usb.affymetrix.com


Mail: Affymetrix UK Ltd.Voyager, Mercury Park, Wycombe Lane, Wooburn Green, High Wycombe HP10 0HH, United Kingdom

To ensure that orders are processed cor-rectly and expediently, please be sure to specify the following: Customer ID number Ship-to address Purchase order or credit card number Product number and name Quantity and size End-user’s name, department, and phone

number VAT identification number (if applicable)

Prices subject to change.

Technical support:In the U.S. and direct:

Phone: 1-888-362-2447 or 216-765-5000 (8:30 a.m. - 5:30 p.m., E.S.T.)

Fax: 800-535-0898 or 216-464-5075


In Europe:

Phone: +44(0)1628 55 2600

Fax: +44(0)1628 55 2675


MSDS requests:On-line: www.usb.affymetrix.com E-mail: [email protected]

Standing orders: Assure timely product delivery on pre-assigned dates. These are multiple, predetermined shipments of the same product or products.

Blanket orders: For added flexibility and convenience when ordering a variety of products over a given period of time.

Bulk orders: This may include large quanti-ties of lot-specific material, custom packag-ing (size, special labeling, etc.), or other unique specifications and requirements.

Shipping information:In the U.S., we use the following shipment methods:

�� All -20°C and 4°C products ship for next-day delivery in most areas.�� All ambient products ship “ground” for 2 to 3 day delivery. If you require next-day delivery for these items, please ask your Customer Service Representative.�� In addition, hazardous products ship via next-day delivery.�� Heavier bulk items can be shipped a number of different ways that best meet your needs and budget.

PRODUCT USES:CAUTION: Products of Affymetrix which are or may be drugs, food additives or diagnostic reagents, as described in the federal food, drug and cosmetic act, are for investigational use only in laboratory research animals or testing in vitro, and are not for drug, new drug, veterinary drug, food, food additive or human use. Unless otherwise indicated, all products are distributed and sold for chemical purposes only, not for drug use or for application to or ingestion by humans or for commercial horticulture use, for pesticide use, for application to or ingestion by animals or for veterinary drug use. All products sold by Affymetrix to Buyer shall be used by qualified professionals only. The burden for safe use and handling of all products sold by Affymetrix to Buyer is entirely the responsibility of Buyer and anyone who purchases the goods from Buyer and uses them. Absence of hazardous warnings does not imply non-toxicity. Any resale, distribution and/or export of products sold by seller to buyer outside the U.S.A. must be strictly in accordance with U.S. law and United Nations regulations.

Please visit www.usb.affymetrix.com/terms for the Affymetrix terms and conditions of sale.

Terms & Conditions

3For bulk or alternate pack sizes, email us at [email protected].

Ordering Information/ISO Certification

The site responsible for manufacturing and distribut-ing the USB-branded products has maintained cer-tification to an ISO standard since 1998. Originally certified to ISO 9000, we are pleased to announce that the site is now certified to ISO 13485:2003. While sharing many similarities with ISO 9001, ISO 13485 demonstrates the site’s preparedness for manufacturing and distributing products which may

be marketed and sold as in vitro diagnostic (IVD) reagents. This is an important change as the site continues to add new capabilities and capacities for Affymetrix.

It is our belief that ISO certification demonstrates our commitment to quality through all segments of the company. Customers can be assured that the employees take quality very seriously. Quality

products and services are obtained through the understanding, participation, and commitment of all employees.

For additional information on the site’s ISO certifica-tion, contact Amy Nasr, Director of Reagent QA and Regulatory Affairs: [email protected], 1-888-362-2447 ext. 5809.

ISO Certification

Affymetrix®, USB®, ExoSAP-IT®, Change-IT™, DeliverX™, FideliTaq™, Genetic Performance Certified™, Hi-Res™, HotStart-IT®, Ligate-IT™,

MagniTaq®, OptiKinase™, Prep2Seq™, PrepEase®, RapidRun™, RubyTaq™, SuperSAP™, Tested User Friendly™, and VeriQuest® are trademarks or

registered trademarks of Affymetrix, Inc. All other trademarks are the property of their respective owners.

Trademarks

Patents & Licenses

DNA Labeling Reagent (DLR) – This product may be covered by one or more of the following patents: U.S. Patent Nos. 6,864,059; 6,965,020; and 7,423,143.

ExoSAP-IT for PCR Product Clean-Up, HT ExoSAP-IT High-Throughput PCR Product Clean-Up, SBE Clean-Up Reagent, SNP-IT Clean-Up Reagent, Sequenase PCR Product Sequencing Kit – These products are covered by patents owned by Affymetrix, including U.S. Patent Nos. 6,379,940 and 6,387,634 and cor-responding patents in Canada and Japan.

ExoSAP-IT for PCR Product Clean-Up, HT ExoSAP-IT High-Throughput PCR Product Clean-Up, SBE Clean-Up Reagent, SNP-IT Clean-Up Reagent, Sequenase PCR Product Sequencing Kit, and the PCR Product Pre-Sequencing Kit – These products or portions thereof are sold under license from GE Healthcare under U.S. Patent Nos. 5,741,676 and 5,756,285.

HotStart-IT Products – These products may be covered by U.S. Patent Nos. 7,700,281; 7,951,534; and 8,404,443, and corresponding foreign patents. These products are also sold under licensing arrangements with Agilent Technologies, Inc. The purchase price of these products includes limited, nontransferable rights under U.S. Patent Nos. 5,605,824, 5,646,019 and 5,773,257 owned by Agilent Technologies to use only this amount of the product to practice the claims in said patents solely for activities of end users within the field of life sci-ence research. Further licensing information may be obtained by contacting the Business Development Department, Agilent Technologies, Inc., 11011 N. Torrey Pines Road, La Jolla, California 92037, USA.

MagniTaq Multiplex PCR Master Mix – Patent pending.

Poly(A) Tail-Length Assay Kit – Patent pending.

PrepEase Genomic DNA Isolation Kit – This product or portions thereof is manufactured under license from Case Western Reserve University and U.S. Pat-ent Number 7,687,254.

RNA Labeling Reagent (RLR) – This product may be covered by one or more of the following patents: U.S. Patent Nos. 6,864,059; 6,965,020; 7,282,327; 7,468,243; and 7,491,818.

Taq DNA Polymerase – Use of this product is covered by one or more of the following U.S. patents and corresponding patent claims outside the U.S.: 5,789,224, 5,618,711, and 6,127,155. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser’s own internal research. No right under any other patent claim, no right to perform any patented method, and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser’s activi-ties for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

SYBR Green Products – Use of these products are covered by one or more of the following US patents and corresponding patent claims outside the US: 5,994,056 and 6,171,785. The purchase of these products includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser’s own internal research. No right under

any other patent claim and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser’s activi-ties for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Direc-tor of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA. This product is also provided under an agreement between Molecular Probes, Inc. and Affymetrix and the manufacture, use, sale or import of this product is subject to one or more U.S. Patents and corresponding international equivalents, owned by Molecular Probes, a Life Technologies Corporation company. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product for real-time PCR conducted by the buyer, for use in research and development, which excludes use in performing testing, analysis or screening services. The buyer cannot sell or otherwise transfer (a) this product (b) its compo-nents or (c) materials made using this product or its components to a third party, whether or not such product or its components are resold for use in research and development, and shall not use this product or its components or materials made using this product or its components for therapeutic or prophylactic purposes. For information on purchas-ing a license to this product for purposes other than set forth above, contact Molecular Probes, Inc., Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA. Tel: (541) 465-8300, Fax: (541) 335-0354.


We deliver custom enzymes and bulk biochemicals with your complete satisfaction as our goal. Our team combines operational flexibility and personalized service with the production resources of a large organization.

Turn to USB for Custom, Bulk, and OEM solutions to fit your specific needs.

Contact us at [email protected] to get started.

USB® has been helping diagnostic, biotech, and pharmaceutical companies succeed in

the life sciences industry for more than 40 years. We can assist you with custom reagent

manufacturing, dispensing, labeling, and kitting of your critical reagent components.

Optimize your use of key staff and limited resources by outsourcing production of your

finished goods or key components to our team.

Custom, Bulk, and OEM


EnzymesLigases, kinases, phosphatases, polymerases, and Sequenase

Convenience ReagentsPre-mixed buffers and stock solutions

PCR ReagentsExoSAP-IT® PCR Cleanup reagent for both standard and high throughput,

SSB binding protein, DNA polymerases, and exo-free klenow

NucleotidesdNTPs, PCR nucleotide mixes, ddNTPs, and NTPs

BiochemicalsAgaroses, acrylamides, amino acids, sugars, substrates, and vitamins

Product PortfolioHere’s a sampling of our more than 3,000 customizable products:

Custom, Bulk, and OEM solutions ■ Custom enzymes, formulated to your special requirements■ Bulk sizes – our enzymes and biochemicals can be packaged into large quantities ■ Special fill sizes/concentrations/formulations of over 3,000 catalog products■ OEM labeling of individual components, or complete kit packaging and labeling with your brand■ General Purpose Reagents (GPR) and IVD reagents & assays to your exacting standards■ Customer-specific quality control assays

The USB team■ Enzymology expertise■ Extensive research, development, and manufacturing experience■ Long term supply agreements and licensing options ■ Experts who will work with you on site or from our reagents center of excellence

ISO 13485 compliant processes■ Rigorous quality system suited for your diagnostic requirements■ Comprehensive documentation■ Change control capabilities ■ We welcome audits of our manufacturing facilities

Custom, Bulk, and OEM

PCR ToolsPCR Amplification

End Point PCR

Long and Accurate PCR

Hot Start PCR

Multiplex PCR

Real-Time PCR (qPCR)

Real-Time Reverse Transcription

PCR (qRT-PCR)

Reverse Transcription PCR (RT-PCR)

PCR Purification

Passive Reference Dyes


PCR Tools | Table of Contents

PCR Product ReviewClear Choices for PCR 8PCR Product Selection Guide 9Instrument Compatibility with VeriQuest qPCR

Master Mixes 10

End Point PCRTaq DNA Polymerase 11Taq PCR Master Mix (2X) 12Taq PCR Kit 12RubyTaq™ DNA Polymerase 13RubyTaq PCR Master Mix (2X) 14

Long and Accurate PCRFideliTaq™ DNA Polymerase 15FideliTaq PCR Master Mix (2X) 16HotStart-IT® FideliTaq DNA Polymerase 17HotStart-IT FideliTaq PCR Master Mix (2X) 18

Hot Start PCRHotStart-IT Taq DNA Polymerase 19HotStart-IT Taq PCR Master Mix (2X) 20HotStart-IT Binding Protein 21

Multiplex PCRMagniTaq® Multiplex PCR Master Mix (2X) 22Promoter Methylation PCR Kit 24HotStart-IT FideliTaq DNA Polymerase 25

Real-Time qPCRVeriQuest® Taq DNA Polymerase 26VeriQuest SYBR® Green qPCR Master Mix (2X) 27VeriQuest SYBR Green qPCR Master Mix with

Fluorescein (2X) 29VeriQuest Fast SYBR Green qPCR Master

Mix (2X) 30VeriQuest Fast SYBR Green qPCR Master Mix

with Fluorescein (2X) 31HotStart-IT SYBR Green qPCR Master Mix (2X) 32HotStart-IT SYBR Green qPCR Master Mix with

UDG (2X) 32VeriQuest Probe qPCR Master Mix (2X) 33VeriQuest Probe qPCR Master Mix, No

Reference Dye (2X) 35VeriQuest Fast Probe qPCR Master Mix (2X) 36VeriQuest Fast Probe qPCR Master Mix, No

Reference Dye (2X) 37HotStart-IT Probe qPCR Master Mix (2X) 38HotStart-IT Probe qPCR Master Mix with

UDG (2X) 38

Real-Time Reverse Transcription PCR (qRT-PCR)First-Strand cDNA Synthesis Kit for Real-Time

PCR 39VeriQuest SYBR Green One-Step qRT-PCR

Master Mix (2X) 40VeriQuest SYBR Green One-Step qRT-PCR

Master Mix with Fluorescein (2X) 41VeriQuest Probe One-Step qRT-PCR Master

Mix (2X) 42VeriQuest Probe One-Step qRT-PCR Master

Mix, No Reference Dye (2X) 43HotStart-IT SYBR Green One-Step qRT-PCR

Master Mix Kit 44HotStart-IT Probe One-Step qRT-PCR Master

Mix Kit 44

Reverse Transcription RT-PCR—One-Step Master MixRT-PCR Master Mix (2X) 45FideliTaq RT-PCR Master Mix (2X) 46

Reverse Transcription RT-PCR—KitsFirst-Strand cDNA Synthesis Kit for Real-Time

PCR 47One-Step RT-PCR Kit 47Two-Step RT-PCR Kit 48

PCR PurificationExoSAP-IT® PCR Product Cleanup 49HT ExoSAP-IT High-Throughput PCR Product

Cleanup 50SBE Cleanup Reagent 52PCR Product Pre-Sequencing Kit 52PrepEase® DNA Cleanup Kits 52PrepEase Gel Extraction Kits 52PrepEase PCR Purification 96-Well Plate Kits

(Ultrafiltration) 53

Passive Reference DyesFluorescein Passive Reference Dye 53ROX™ Passive Reference Dye 53


End Point PCR

Standard Taq DNA Polymerase [71160] Taq PCR Master Mix (2X) [71162] RubyTaq DNA Polymerase [71190] RubyTaq PCR Master Mix (2X) [71191]

Long and Accurate PCR FideliTaq DNA Polymerase [71180] FideliTaq PCR Master Mix (2X) [71182] HotStart-IT FideliTaq DNA Polymerase [71155] HotStart-IT FideliTaq PCR Master Mix (2X) [71156]

Hot Start HotStart-IT Taq DNA Polymerase [71195] HotStart-IT Taq PCR Master Mix (2X) [71196] HotStart-IT Binding Protein [71194]

Multiplex Promoter Methylation PCR Kit [MP1100] MagniTaq Multiplex PCR Master Mix (2X) [71199]

PCR Tools | PCR Product Review

Clear Choices for PCR

Real-Time qPCR

Components & Kits First-Strand cDNA Synthesis Kit for Real-Time PCR [75780] VeriQuest Taq DNA Polymerase (2X) [71170]

SYBR VeriQuest SYBR Green qPCR Master Mix (2X) [75600] VeriQuest SYBR Green qPCR Master Mix with Fluorescein

(2X) [75665] VeriQuest Fast SYBR Green qPCR Master Mix (2X) [75690] VeriQuest Fast SYBR Green qPCR Master Mix with

Fluorescein (2X) [75675] HotStart-IT SYBR Green qPCR Master Mix (2X) [75762] HotStart-IT SYBR Green qPCR Master Mix with UDG (2X)

[75760]

Probe VeriQuest Probe qPCR Master Mix (2X) [75650] VeriQuest Probe qPCR Master Mix, No Reference Dye (2X)

[75660] VeriQuest Fast Probe qPCR Master Mix (2X) [75680] VeriQuest Fast Probe qPCR Master Mix, No Reference Dye

(2X) [75685] HotStart-IT Probe qPCR Master Mix (2X) [75766] HotStart-IT Probe qPCR Master Mix with UDG (2X) [75764]

Real-Time Reverse Transcription PCR (qRT-PCR)

One-Step Master Mixes VeriQuest SYBR Green One-Step qRT-PCR Master Mix (2X)

[75705] VeriQuest SYBR Green One-Step qRT-PCR Master Mix with

Fluorescein (2X) [75715] VeriQuest Probe One-Step qRT-PCR Master Mix (2X)

[75700] VeriQuest Probe One-Step qRT-PCR Master Mix, No

Reference Dye (2X) [75710] HotStart-IT SYBR Green One-Step qRT-PCR Master Mix Kit

[75770] HotStart-IT Probe One-Step qRT-PCR Master Mix Kit

[75772]

Kits First-Strand cDNA Synthesis Kit for Real-Time PCR [75780]

Reverse Transcription RT-PCR

One-Step Master Mix RT-PCR Master Mix [2X] [78370] FideliTaq RT-PCR Master Mix [2X] [71185]

Kits One-Step RT-PCR Kit [78350] Two-Step RT-PCR Kit [78355]

PCR Components

Nucleotides 10 mM PCR Nucleotide Mix [77212] 25 mM PCR Nucleotide Mix [77119] 10 mM PCR Nucleotide Mix with dUTP [77330]

Reagents 5M Betaine [77507] Nuclease-Free Water [71786]



PCR Product Selection Guide

End

Point

PCR

Hot St

art P

CR

Long

and

Accur

ate P

CR

Mult

iplex

PCR

Fast

PCR

Trac

king

Dyes P

CR

Mas

ter M

ix Fo

rmat

Stan

d-alo

ne En

zym

e

RT-P

CR

Real-

time P

CR: SY

BR

Real-

time P

CR: Pr

obe

One-S

tep

qRT-

PCR:

SYBR

One-S

tep

qRT-

PCR:

Prob

e

Taq DNA Polymerase [71160] � �Taq PCR Master Mix (2X) [71162] � �RubyTaq™ DNA Polymerase [71190] � � �RubyTaq PCR Master Mix (2X) [71191] � � �FideliTaq™ DNA Polymerase [71180] � � �FideliTaq PCR Master Mix (2X) [71182] � � �VeriQuest® Taq DNA Polymerase [71170] � � �HotStart-IT® Taq DNA Polymerase [71195] � � �HotStart-IT Taq Master Mix (2X) [71196] � � �HotStart-IT FideliTaq DNA Polymerase [71155] � � � �HotStart-IT FideliTaq PCR Master Mix (2X) [71156] � � � �

AMV Reverse Transcriptase [70041] � �M-MLV Reverse Transcriptase [78306] � �Tth DNA Polymerase [70052] � �RT-PCR Master Mix (2X) [78370] � �FideliTaq RT-PCR Master Mix (2X) [71185] � �

First-Strand cDNA Synthesis Kit for Real-Time PCR [75780] � � � �VeriQuest SYBR® Green qPCR Master Mix (2X) [75600] � � �VeriQuest SYBR Green qPCR Master Mix with Fluorescein (2X) [75665] � � �VeriQuest Probe qPCR Master Mix (2X) [75650] � � �VeriQuest Probe qPCR Master Mix, No Reference Dye (2X) [75660] � � �VeriQuest Fast SYBR Green qPCR Master Mix (2X) [75690] � � � �VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein (2X) [75675] � � � �VeriQuest Fast Probe qPCR Master Mix (2X) [75680] � � � �VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X) [75685] � � � �VeriQuest SYBR Green One-Step qRT-PCR Master Mix (2X) [75705] � � �VeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein (2X) [75715] � � �VeriQuest Probe One-Step qRT-PCR Master Mix (2X) [75700] � � �VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye (2X) [75710] � � �HotStart-IT SYBR Green qPCR Master Mix (2X) [75762] � � �HotStart-IT SYBR Green qPCR Master Mix with UDG (2X) [75760] � � �HotStart-IT Probe qPCR Master Mix (2X) [75766] � � �HotStart-IT Probe qPCR Master Mix with UDG (2X) [75764] � � �

MagniTaq® Multiplex PCR Master Mix [71199] � � � �

Let our simple chart below help you find the optimal reagent for your research needs. Navigate our PCR product portfolio by application to view your options.



Instrument Compatibility with VeriQuest qPCR Master Mixes

Use this chart to determine which of our VeriQuest qPCR master mixes we recommend for your machine. Simply find your thermo cycler across the top row and then locate which of our master mixes are optimized for that instrument.

VeriQuest Products

Enzyme

71170 VeriQuest Taq DNA Polymerase See page 26

SYBR Green Mixes

75600 VeriQuest SYBR Green qPCR Master Mix (2X) See page 27

75665 VeriQuest SYBR Green qPCR Master Mix with Fluorescein (2X) See page 29

Probe Mixes

75650 VeriQuest Probe qPCR Master Mix (2X) See page 33

75660 VeriQuest Probe qPCR Master Mix, No Reference Dye (2X) See page 35

Fast SYBR Green Mixes

75690 VeriQuest Fast SYBR Green qPCR Master Mix (2X) See page 30

75675 VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein (2X) See page 31

Fast Probe Mixes

75680 VeriQuest Fast Probe qPCR Master Mix (2X) See page 36

75685 VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X) See page 37

One-Step qRT-PCR SYBR Green Mixes

75705 VeriQuest SYBR Green One-Step qRT-PCR Master Mix (2X) See page 40

75715 VeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein (2X) See page 41

One-Step qRT-PCR Probe Mixes

75700 VeriQuest Probe One-Step qRT-PCR Master Mix (2X) See page 42

75710 VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye (2X) See page 43

SYBR Green Mix ABI Pri

sm® 70

00

ABI Pri

sm 77

00

ABI Pri

sm 79

00 HT*

ABI 57

00

ABI 73

00

ABI 75

00*

ABI Ste

pOne

™/St

epOne

Plus™

*

ABI ViiA

™ 7

Bio-Ra

d iCycl

er iQ

® /iQ5

Bio-Ra

d iCycl

er MyiQ

™

Bio-Ra

d CFX

96™

/CFX38

4™

Bio-Ra

d Opti

con2

™

Bio-Ra

d Chro

mo4™

Cephe

id Sm

art Cycl

er®

Corbett

Rotor

-Gen

e™

Eppe

ndorf

Mast

ercycl

er® ep

realp

lex

Fluidi

gm Bi

oMark

™

Illum

ina Ec

o™*/H

elixis

Pixo

™

Qiagen

Rotor

-Gen

e™ Q

*

Roch

e Ligh

tCycler

® 1.0,

1.5, 2

.0*

Roch

e Ligh

tCycler

480/1

536*

Strata

gene

Mx3

000P

™*

Strata

gene

Mx3

005P

™*

Strata

gene

Mx4

000™

TaKaR

a TP-8

00™

VeriQuest® SYBR® Green qPCR Master Mix (2X) [75600]

VeriQuest SYBR Green qPCR Master Mix with Fluorescein (2X) [75665]

Probe Mix

VeriQuest Probe qPCR Master Mix (2X) [75650]

VeriQuest Probe qPCR Master Mix, No Reference Dye (2X) [75660]

Fast SYBR Green Mix

VeriQuest Fast SYBR Green qPCR Master Mix (2X) [75690]

VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein (2X) [75675]

Fast Probe Mix

VeriQuest Fast Probe qPCR Master Mix (2X) [75680]

VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X) [75685]

One-Step qRT-PCR SYBR Green Mix

VeriQuest SYBR Green One-Step qRT-PCR Master Mix (2X) [75705]

VeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein (2X) [75715]

One-Step qRT-PCR Probe Mix

VeriQuest Probe One-Step qRT-PCR Master Mix (2X) [75700]

VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye (2X) [75710]

* Instruments can be set to fast mode cycling when using fast mode master mixes. Indicates preferred kit for this cycler

Indicates maybe also used for this cycler


PCR Tools | End-Point PCR

71160 50 units 250 units

1,000 units 5,000 unitsCustoms by request

Source:E. coli strain expressing a full length, unmodified clone of Taq DNA Polymerase from Thermus aquaticus(1-3).

Description:Taq DNA Polymerase is a thermostable enzyme which has a highly processive 5'→3' polymerase activity and a 5'→3' exonuclease activity (1). Taq enzyme is functionally tested and has no detectable contaminating endonucleases. Taq DNA Polymerase withstands repeated incubations at 95°C without a significant decrease in enzyme activity, and is suitable for routine PCR. Taq DNA Polymerase is sup-plied with 10X PCR Reaction Buffer plus a separate tube of 25 mM MgCl2 for optimizing PCR conditions. Each lot is tested for product yield and length in PCR amplification.

Applications:�� Routine PCR amplification�� Suitable for real-time qPCR

Properties:Activator: Mg2+

Purity:Free from detectable non-specific nucleases.

Storage buffer:20 mM Tris-HCl, pH 8.5, 1 mM DTT, 0.1 mM EDTA, 100 mM KCI, 50% glycerol, stabilizers.

Unit definition:One unit incorporates 10 nmol of total nucleotides into acid-insoluble material in 30 minutes at 74°C in a total volume of 50 µl.

Concentration:5 units/µl

Assay conditions:The reaction mixture (50 µl) contains 25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM 2-mercaptoethanol, 200 µM dNTPs, 250 µg/ml activated salmon sperm DNA, and Taq DNA Polym-erase. After incubation at 74°C for 10 minutes, acid insoluble material is determined.

Functional test:Tested in a standard PCR.

Functionally tested 10X PCR Reaction Buffer (included):100 mM Tris-HCl (pH 8.6), 500 mM KCl, 15 mM MgCl2Functionally tested MgCl2 (included):25 mM solution

Shipping and storage:Shipped on dry ice. Store at -20°C.

References:1. Innis, M. A., Myambo, K. B., Gelfand, D. H. and

Brow, M. A. (1988) Proc. Natl. Acad. Sci. 85, 9436-9440.

2. Lawyer, F. C., Stoffel, S., Saiki, R. K., Myambo, K., Drummond, R. and Gelfand, D. H. (1989) J. Biol. Chem. 264, 6427-6437.

3. Innis, M. A. and Gefland, D. H. (1990) PCR Protocols: A Guide to Methods and Applications, Academic Press.

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ExoSAP-IT PCR Product Cleanup78250 20 reactions78200 100 reactions78201 500 reactions78202 2,000 reactions78205 5,000 reactions

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PCR Nucleotide Mix, 10 mM Solution(10 mM each dATP, dCTP, dGTP, dTTP)77212 500 µl

PCR Nucleotide Mix, 25 mM Solution77119 500 µl

PCR Nucleotide Mix with dUTP, 10 mM Solution77330 500 µl

Water, Nuclease-Free71786 10 x 1 ml 100 ml 500 ml 1 L 5 L

Taq PCR Kit71161 100 reactions

Taq DNA Polymerase

Fig. 1. DNA fragments ranging from 1 kb to 6 kb amplified from λ DNA.

0.5 kb –

1 kb –

2 kb –

5 kb –

12 kb –

M 1 2 3 4 5 6 kbp


71162 100 reactionsCustoms by request

Taq PCR Master Mix is supplied as a convenient 2X pre-mixed formulation containing Taq DNA Polymerase, Ultrapure nucleotides, and reaction buffer optimized for a wide variety of PCR applica-tions. Taq PCR Master Mix provides robust and reliable performance in PCR amplification. Since the mix is prepared from high quality reagents and is thoroughly QC tested, experimental variability is significantly reduced.

Applications:�� Routine PCR amplification�� Suitable for real-time qPCR�� High-throughput PCR

ConvenientThe mix saves time and reduces potential contami-nation errors by eliminating several pipetting steps. For a 50 µl reaction, simply add 25 µl of Taq PCR Master Mix to primers, DNA template, and PCR-Qualified H2O.

StableThe mix withstands repeated freeze-thaw cycles with no observed decrease in performance (Fig. 1). It is also stable at 4°C for extended periods of time. In fact, rigorous stability studies have been done at Affymetrix showing the mix to be stable at room temperature (Fig. 2).

ScaleableThe end-user may set-up 10 µl, 25 µl, 50 µl, or 100 µl reactions.

Ideal for high-throughputTaq PCR Master Mix is ideal for high-throughput ap-plications such as screening transformants by colony PCR (Fig. 3) or transgenic studies.

Sensitive and robustThe mix may be used to amplify PCR products from low-copy genomic DNA targets as well as low to moderately abundant cDNAs (Figs. 1-3). Excellent results are also obtained with long PCR products (Fig. 4).

Functional test:Functionally tested in PCR according to the standard Affymetrix PCR protocol. This protocol is based upon the amplification of a 500 bp DNA sequence.

Taq PCR Master Mix Formulation (2X):20 mM Tris-HCl (pH 8.6), 100 mM KCl, 3 mM MgCl2, 0.4 mM dNTPs (dATP, dCTP, dGTP, dTTP), 50 units/ml Taq DNA Polymerase, stabilizers.

Components:4 x 625 µl (100 x 50 µl reactions or 200 x 25 µl reactions)

Shipping and storage:Shipped on dry ice. Store at -20°C. Mix well prior to use.


Taq PCR Master Mix (2X)

Fig. 1. Freeze-thaw stability of Taq PCR Master Mix. Amplification of a segment of the single-copy numb gene from 50 pg human genomic DNA. This represents about 30 total target copies. Following 25 freeze-thaw cycles, no significant effect on performance was observed.

Fig. 2. Elevated temperature stability. Taq PCR Master Mix was stored at 4°C and room temper-ature for two months and its performance was compared against a freshly prepared mix. Segments of the single-copy p53 gene (left) and numb gene (right) were amplified from 50 pg human genomic DNA. (This represents about 30 total target copies.) Under these storage conditions, PCR amplifica-tion was not significantly affected.

Fig. 3. Colony PCR. Amplification of a segment of the T4 DNA Polymerase gene overexpressed in several different E. coli M5248 colonies.

Fig. 4. Amplification of targets from genomic DNA. Segments of the single-copy NRAGE gene (a) and numb gene (b) were amplified from 50 pg and 50 ng of human genomic DNA respectively.

M 0 15 25Freeze thaws

0.4 kb –

1.0 kb –

← numb (455 bp)

M Fresh 4°C RT M Fresh 4°C RT

p53 (247 bp) →

0.5 kb –

0.1 kb –

← numb (455 bp)

M

← 574 bp1.0 kb –

0.5 kb –

M

NRAGE (1369 bp) →

a

2.0 kb –

1.0 kb –

← numb (4.6 kb)

M

b

5.0 kb –

3.0 kb –

71161 100 reactions (100 µl/rctn)

The Taq PCR Kit includes the necessary reagents to amplify DNA from a wide variety of templates. All components, including Ultrapure PCR Qualified Water and Ultrapure PCR Nucleotide Mix, are

functionally tested in standard PCR to ensure reliable results. The kit also includes two PCR buffers for standard reactions, one with and one without MgCl2, and a separate tube of 25 mM magnesium chloride for optimizing PCR conditions.

Kit components:Taq DNA Polymerase, 250 units10 mM PCR Nucleotide Mix, Ultrapure10X PCR Buffer with MgCl210X PCR Buffer without MgCl225 mM MgCl2PCR-Qualified Water, Ultrapure

Taq PCR Kit





Source:E. coli strain expressing a clone of Taq DNA Polym-erase from Thermus aquaticus(1-3).

Description:RubyTaq DNA Polymerase contains high-quality Taq DNA Polymerase with two inert dyes that serve as tracking dyes in gel electrophoresis. This greatly simplifies analysis of PCR reactions because no load-ing dyes or compounds that increase sample density need to be added prior to gel electrophoresis. During gel electrophoresis, RubyTaq Polymerase separates into 2 colors, magenta and yellow, for easy visualiza-tion (Fig. 1). The magenta dye runs in a range be-tween 500 bp (2% gels) and 1,500 bp (0.8% gels), and the yellow dye runs at less than 10 bp.

Applications:�� Routine PCR amplification�� TA-vector cloning�� Amplification prior to in vitro transcription

Easy-to-see, easy-to-load�� Load directly to gel from thermocycler�� No additional loading buffer needed�� RubyTaq separates into 2 colors – magenta and yellow – during gel electrophoresis.

Because the dyes are inert during PCR, RubyTaq DNA Polymerase provides the same outstanding performance as standard Taq DNA Polymerase (Fig. 2). In addition, the dyes do not affect the migration rates of DNA and have no effect on the following procedures:�� PCR with co-solvents (e.g., betaine and DMSO)�� Restriction digestion�� Automated or manual DNA sequencing�� Ligation-based or topoisomerase-based (TOPO®) TA cloning�� ExoSAP-IT PCR Cleanup

If the dyes must be removed prior to another ap-plication (e.g., in vitro transcription), use phenol/chloroform extraction and ethanol precipitation, or the PrepEase Sequencing Dye Cleanup Kit (PN 78776/77).



Storage buffer:20 mM Tris-HCl, pH 8.5, 1 mM DTT, 0.1 mM EDTA, 50 mM KCI, 50% glycerol, inert dyes, and stabilizers.


Concentration:1 unit/µl

Assay conditions:The reaction mixture (50 µl) contains 25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM 2-mercaptoethanol, 200 µM dNTPs, 250 µg/ml acti-vated salmon sperm DNA, and cloned RubyTaq DNA Polymerase. After incubation at 74°C for 10 minutes, acid-insoluble material is determined.

Functional test:Functionally tested for PCR.

Functionally tested 10X PCR Reaction Buffer without MgCl2 (included):100 mM Tris-HCl (pH 8.6) and 500 mM KCl

Functionally tested MgCl2 (included):25 mM solution





3. Innis, M. A. and Gefland, D. H. (1990) PCR Protocols: A Guide to Methods and Applications, Academic Press.

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TAE Buffer, 10X Solution75904 1 L 5 L

TBE Buffer, 5X Solution75891 1 L 5 L


RubyTaq DNA Polymerase

Fig. 2. Performance of RubyTaq DNA Polymerase versus Taq DNA Polymerase. Single-copy targets were amplified from the indi-cated amounts of human genomic DNA. For the 455 bp numb ampli-con, 50 pg of template represents about 30 total target copies in the reaction. T is Taq DNA Polymerase, R is RubyTaq DNA Polymerase, and M is the DNA marker.

M T R T R T R

5.0 kb – ← 4.6 kb numb (50 ng)← 1.55 kb NRAGE (10 ng)

← 455 bp numb (50 pg)

1.0 kb –

100 bp –

Fig. 1. RubyTaq DNA Polymerase visualization during gel electrophoresis. RubyTaq Polymerase in PCR does not require the use of additional loading buffer. Simply load onto an agarose gel directly after cycling. During electrophoresis, RubyTaq separates into 2 colors, magenta (runs between 500 bp [2% gels] and 1,500 bp [0.8% gels]) and yellow (runs less than 10 bp). The indicated volumes of RubyTaq DNA Polymerase were run on a 1% TAE agarose gel.


71191 100 reactions 500 reactionsCustoms by request

RubyTaq PCR Master Mix is supplied as a convenient 2X pre-mixed formulation containing Taq DNA Polymerase, Ultrapure nucleotides, reaction buffer, and two inert dyes optimized for a wide variety of PCR applications. RubyTaq enzyme greatly simplifies analysis of PCR reactions because no loading dyes or compounds that increase sample density need to be added prior to gel electrophoresis. During gel elec-trophoresis, RubyTaq PCR Master Mix separates into 2 colors, magenta and yellow, for easy visualization. The magenta dye runs in a range between 500 bp (2% gels) and 1,500 bp (0.8% gels) and the yellow dye runs at less than 10 bp.

RubyTaq PCR Master Mix has been designed to streamline high-throughput PCR applications and provides for robust and reliable performance. Since the mix is prepared from high quality reagents and is thoroughly QC tested, experimental variability is significantly reduced.

Applications:�� Routine PCR amplification�� TA vector cloning�� Amplification prior to in vitro transcription�� High-throughput PCR

Easy-to-see, easy-to-load�� Load directly to gel from thermocycler�� No additional loading buffer needed�� RubyTaq separates into 2 colors – magenta and yellow – during gel electrophoresis.

Ultimate convenienceThe mix saves time and reduces potential contami-nation errors by eliminating several pipetting steps. For a 50 µl reaction, simply add 25 µl of RubyTaq PCR Master Mix to primers, DNA template, and PCR-Qualified H2O. Reactions can be easily performed in 10 µl, 25 µl, 50 µl, or 100 µl volumes. In addition, PCR reactions are loaded directly onto agarose gels without adding any loading dyes or buffers.

StableThe mix withstands repeated freeze-thaw cycles and extended periods at room temperature with no observed decrease in performance (Fig. 1).

Ideal for high-throughputRubyTaq PCR Master Mix is ideal for high-throughput applications such as:�� Screening transformants by colony PCR�� Transgenic genotyping�� Ligation-based or topoisomerase-based (TOPO) TA cloning

The mix may be used to amplify low-copy targets from complex templates and can also amplify a variety of product lengths.

Functional test:Functionally tested for PCR according to the stan-dard Affymetrix PCR protocol. This protocol is based upon the amplification of a 500 bp DNA sequence.

RubyTaq PCR Master Mix Formulation (2X):20 mM Tris-HCl (pH 8.6), 100 mM KCl, 4 mM MgCl2, 0.4 mM dNTPs (dATP, dCTP, dGTP, dTTP), 80 units/ml Taq DNA Polymerase, inert dyes, stabilizers.

Components:4 x 625 µl RubyTaq PCR Master Mix, 2X

(100 x 50 µl reactions or 200 x 25 µl reactions)


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RubyTaq PCR Master Mix (2X)


M

← 455 bp numb (50 pg)1.0 kb –

100 bp –

-20°CF/T RT

Fig. 1. Stability of RubyTaq PCR Master Mix. RubyTaq PCR Master Mix stability was assessed by comparing a reference mix stored at -20°C versus a sample subjected to 15 freeze-thaw cycles (F/T) and a sample stored at room temperature for one month (RT). The single-copy numb gene was amplified from 50 pg of human genomic DNA which represents about 30 total target copies in the reaction. Under these test conditions, performance of the master mix is not significantly affected.



1,000 units 5 x 250 units 5,000 unitsCustoms by request

Source:Recombinant proteins expressed in E. coli.

Description:FideliTaq DNA Polymerase combines high quality recombinant Taq DNA Polymerase with a high-fidelity, proofreading polymerase. This enzyme blend has the 5'→3' exonuclease activity of Taq DNA Polymerase as well as the 3'→5' exonuclease activity of the proofreading polymerase. FideliTaq DNA Polymerase increases amplification fidelity up to 6 times over Taq DNA Polymerase alone and allows for amplification of longer product sizes(1-4) (Fig. 1). The enzyme blend generates PCR products whose ends are compatible with either blunt-end or TA cloning procedures(5) with A-tailed ends favored over blunt ends in an approximately 3 to 1 ratio.

Applications:�� PCR-mediated cloning�� Long and accurate PCR amplification


Purity:Free of detectable non-specific nucleases.


Unit definition:One unit incorporates 10 nmol of total nucleotides into acid-insoluble material in 30 minutes at 74°C in a total volume of 50 µl(6-7).


Assay conditions:The reaction mixture contains 25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM β-ME, 200 µM dNTPs, 250 µg/ml activated salmon sperm DNA, and FideliTaq DNA Polymerase. Following incubation at 74°C for 10 minutes, acid-insoluble material is determined.

Functional test:Functionally tested in PCR to amplify a 20.7 kb product.

Functionally tested 10X PCR Reaction Buffer (included):100 mM Tris-HCl (pH 8.6), 500 mM KCl, 15 mM MgCl2Functionally tested MgCl2 (included):25 mM solution


References:1. Barnes, W. M. (1994) Proc. Natl. Acad. Sci. USA

91, 2216-2220.2. Cheng, S., Fockler, C., Barnes, W. M., and

Higuchi, R. (1994) Proc. Natl. Acad. Sci. USA 91, 5695-5699.

3. Barnes, W. M. (1992) Gene 112, 29-35.4. Cline, J., Braman, J. C., and Hogrefe, H. H. (1996)

Nucl. Acids Res. 24, 3546-3551.5. Magnuson, V. L., Ally, D. S., Nylund, S. J.,

Karanjawala, Z. E., Rayman, J. B., Knapp, J. I., Lowe, A. L., Ghosh, S., and Collins, F. S. (1996) BioTechniques 4, 700-709.

6. Innis, M. A., Myambo, K. B., Gelfand, D. H. and Brow, M. A. (1988) Proc. Natl. Acad. Sci. 85, 9436-9440.


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PCR Markers, 50 - 2000 bp76710 250 µl



FideliTaq DNA Polymerase

PCR Tools | Long and Accurate PCR

Fig. 1. Amplification of diverse targets by FideliTaq DNA Polymerase. Target (source): Lane 1: Numb, 455 bp (10 ng human genomic DNA); Lane 2: NRAGE, 1.55 kb (10 ng human genomic DNA); Lane 3: Numb, 4.6 kb (10 ng human genomic DNA); Lane 4: Lambda target, 20.7 kb (10 ng lambda DNA). A total of 25 cycles was used for lambda target and 35 cycles for human genomic DNA targets. A final concentration of 1.5 mM MgCl2 was used in all reactions.

M 1 2 3 4

12.0 kb –

3.0 kb –

1.65 kb –

500 bp –



71182 100 reactionsCustoms by request

FideliTaq PCR Master Mix (2X) combines Taq DNA Polymerase, a high-fidelity, proofreading polym-erase and Ultrapure nucleotides in a proprietary buffer formulation. This ready-to-use mix increases amplification fidelity up to 6 times over Taq DNA Polymerase alone and allows for amplification of longer product sizes(1-4). The enzyme blend generates PCR products whose ends are compatible with either blunt-end or TA cloning procedures(5) with A-tailed ends favored over blunt ends in an approximately 3 to 1 ratio. FideliTaq PCR Master Mix provides robust and reliable performance for PCR applications in which high-fidelity or longer product sizes are re-quired. Since the mix is pre-formulated, experimental variability is significantly reduced.

Applications:�� PCR-mediated cloning�� Long and accurate PCR amplification�� High-throughput PCR

ConvenientThe mix saves time and reduces potential contami-nation errors by eliminating several pipetting steps. For a 50 µl reaction, simply add 25 µl of FideliTaq PCR Master Mix to primers, DNA template, and PCR-Qualified H2O. Reactions can be easily performed in 10 µl, 25 µl, 50 µl or 100 µl volumes.

High fidelityFideliTaq DNA Polymerase gives up to 6-fold higher fidelity than Taq DNA Polymerase, ideal for cloning and microarray applications.

Increase product size and yieldAmplify short and long PCR products from complex DNA templates with little or no optimization (Fig. 1). For PCR products greater than 2 kb, yields are greatly increased with FideliTaq DNA Polymerase and an enhanced buffer system.

Improve specificity and sensitivityAmplify PCR products with low background and from low-copy targets, essential for demanding genomic and cDNA applications with limited sample material (Fig. 2).

StableThe mix withstands repeated freeze-thaw cycles with no observed decrease in performance (Fig. 3).

Functional test:Functionally tested in PCR to amplify a 20.7 kb product.

FideliTaq PCR Master Mix Formulation (2X):50 mM Tris, pH 8.8, 100 mM KCl, 3 mM MgCl2, 0.4 mM dNTPs (dATP, dCTP, dGTP, dTTP), 50 units/ml FideliTaq DNA Polymerase, stabilizers.

Components:4 x 625 µl FideliTaq PCR Master Mix, 2X

(100 x 50 µl reactions or 200 x 25 µl reactions)



91, 2216-2220.2. Cheng, S., Fockler, C., Barnes, W. M., and





FideliTaq PCR Master Mix (2X)

Fig. 1. Range of targets amplified with FideliTaq PCR Master Mix. Single-copy NRAGE, 1.55 kb (Lane 1) and Numb, 7.7 kb (Lane 2) were amplified from 1 ng human genomic DNA. Single-copy β-globin, 23.0 kb (Lane 3) was amplified from 100 ng human genomic DNA. Both the 20.7 kb (Lane 4) and 35.0 kb (Lane 5) lambda targets were amplified from 1 ng lambda DNA. No magnesium optimization was required, as 1.5 mM final magnesium concentration was used in all reactions.

M 1 2 3 4 5

12.0 kb –

7.0 kb –

1.65 kb –

500 bp –

Fig. 2. Sensitivity of FideliTaq PCR Master Mix. The single-copy Numb gene was amplified from the indicated amounts of human genomic DNA. Approximately one mammalian cell is represented by 3 pg of genomic DNA.

M 300 30 3 pg

650 bp –500 bp –400 bp –

Fig. 3. Freeze-thaw stability of FideliTaq PCR Master Mix. The master mix was subjected to 15 freeze-thaw cycles alternating between dry ice and room temperature. Following freeze-thaw cycles (F/T), the mix was compared to control mix (C) before treatment. Both short and long targets are shown to demonstrate the robust nature of the mix. Single-copy Numb, 455 bp (Lane 1) was amplified from 30 pg human genomic DNA. β-globin 23 kb target (Lane 2) was amplified from 100 ng human genomic DNA. Lambda 35 kb target (Lane 3) was amplified from 1 ng lambda DNA.

1 1 2 2 3 3M C F/T C F/T C F/T

12.0 kb –

500 bp –





Source:All proteins are recombinant versions expressed in E. coli.

Description:HotStart-IT FideliTaq DNA Polymerase combines a novel hot start method developed at Affymetrix with the long and accurate amplification properties of FideliTaq DNA Polymerase. The hot start method, called primer sequestration, uses a binding protein to reduce or eliminate non-specific primer-extension products which may be generated at lower tempera-tures during assembly of PCR reactions. Following the initial denaturation step during PCR, the binding protein is inactivated and the primers are free to participate in the amplification reaction. This novel method enhances many complex PCR reactions by increasing specificity and yield (Fig. 1) as well as sensitivity (Fig. 2).

FideliTaq DNA Polymerase combines Taq DNA Polym-erase with a high-fidelity, proofreading polymerase. This enzyme blend has the 5'→3' exonuclease activity of Taq DNA Polymerase as well as the 3'→5' exonuclease activity of the proofreading enzyme. FideliTaq DNA Polymerase increases fidelity up to 6-fold over Taq DNA Polymerase alone and allows for amplification of longer products(1-4) (Fig. 3). The enzyme blend generates PCR products whose ends are compatible with either blunt-end or TA cloning procedures(5), with A-tailed ends favored over blunt ends in an approximately 3 to 1 ratio.

Applications:�� High-specificity and high-sensitivity PCR amplification�� Extremely long PCR amplification�� PCR-mediated cloning

Advantages:�� Room temperature reaction set-up.�� Minimizes amplification of non-specific products and primer-dimers (Fig. 1).�� High specificity and sensitivity (Fig. 2)�� Generates long PCR amplicons (Fig. 3)�� Up to 6-fold higher fidelity than Taq polymerase



Storage buffer:20 mM Tris-HCl (pH 8.5), 1 mM DTT, 0.1 mM EDTA, 200 mM KCI, 50% glycerol, and stabilizers.


Concentration:2.5 units/µl

Assay conditions:The reaction mixture contains 25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM 2-mercaptoethanol, 200 µM dNTPs, 250 µg/ml activated salmon sperm DNA, and Taq DNA Polymerase. Following incubation at 74°C for 10 minutes, acid-insoluble material is determined.

Functional tests:PCR with HotStart-IT FideliTaq DNA Polymerase shifts production of primer-dimers to a specific target of 306 bp from 1 ng of human genomic DNA rela-tive to FideliTaq DNA Polymerase.

PCR with HotStart-IT FideliTaq DNA Polymerase generates a 20.7 kb product.

Functionally tested 10X PCR Reaction Buffer (included): 100 mM Tris-HCl (pH 8.6), 500 mM KCl, 15 mM MgCl2Functionally tested MgCl2 (included):25 mM solution



91, 2216-2220. 2. Cheng, S., Fockler, C., Barnes, W. M., and


3. Barnes, W. M. (1992) Gene 112, 29-35. 4. Cline, J., Braman, J. C., and Hogrefe, H. H. (1996)

Nucl. Acids Res. 24, 3546-3551. 5. Magnuson, V. L., Ally, D. S., Nylund, S. J.,




HotStart-IT FideliTaq DNA Polymerase

Fig. 3. Extreme amplicon length using HotStart-IT FideliTaq DNA Polymerase. Two long PCR products of 20.7 kb and 35 kb were amplified from lambda DNA using 1 ng and 10 ng of DNA respectively. Cycling conditions consisted of a short initial denatur-ation phase at 95°C for 20 seconds and 25 cycles of 95°C for 5 sec-onds and 68°C for 20 minutes.

Fig. 2. Sensitivity of HotStart-IT FideliTaq DNA Polymerase. A 455 bp fragment of the single-copy numb gene was amplified from the indicated amounts of human genomic DNA. The polymerase is extremely sensitive as amplification can be achieved from approxi-mately one human cell.

500 bp –

100 bp –

3 pg (1 cell)

30 pg

– numb, 455 bp

300 pg

500 bp –

100 bp –

no HotStart

HotStart

– numb, 306 bp

– primer-dimers

12 kb –

5 kb –

20.7 kb35 kb

Fig. 1. Increased specificity of HotStart-IT FideliTaq DNA Polymerase. A 306 bp fragment of the single-copy numb gene was amplified from 1 ng of human genomic DNA with and without HotStart-IT technology. The primers in this assay were designed with 3 bases of overlap at their 3'-ends to favor primer-dimer formation during reaction set-up at room temperature. Results demonstrate a shift from mainly primer-dimers to the desired product when HotStart-IT is used.


71156 25 reactions 100 reactions 500 reactionsCustoms by request

HotStart-IT FideliTaq Master Mix combines a novel hot start method developed at Affymetrix with the long and accurate amplification properties of FideliTaq DNA Polymerase. The hot start method, called primer sequestration, uses a binding protein to reduce or eliminate non-specific primer-extension products which may be generated at lower temperatures during assembly of PCR reactions. Following the initial denaturation step during PCR, the binding protein is inactivated and the primers are free to participate in the amplification reaction. This novel method enhances many complex PCR reactions by increasing specificity and yield (Fig. 1) as well as sensitivity (Fig. 2).

HotStart-IT FideliTaq Master Mix combines HotStart-IT FideliTaq DNA Polymerase and Ultrapure nucleotides in a proprietary reaction buffer. Simply add template, primers, and water, and the reactions are ready for cycling. The mix increases amplification fidelity up to 6-fold over Taq DNA Polymerase alone and generates much longer PCR products(1-4) (Fig. 3). PCR products have ends which are compatible with either blunt-end or TA cloning procedures(5) with A-tailed ends favored over blunt ends in an approximately 3 to 1 ratio.

Applications:�� High-specificity and high-sensitivity PCR amplification�� Extremely long PCR amplification�� PCR-mediated cloning�� High-throughput PCR

ConvenientSave time and reduce potential contamination errors by eliminating several pipetting steps. For a 50 µl reaction, simply add 25 µl of HotStart-IT FideliTaq Master Mix to primers, DNA template, and PCR-Qualified H2O. Reactions can be tailored from 10 µl to 100 µl volumes.

Room temperature reaction assembly is possible because of the hot start feature.

Novel hot start technologySince the polymerase is not chemically-inactivated, no extensive heat-activation step is necessary which reduces damage to precious DNA samples.

Higher specificity and sensitivityMinimize amplification of non-specific products and primer-dimers (Fig. 1). Amplify PCR products with low background and from low-copy targets, essential for demanding genomic and cDNA applications with limited sample material (Fig. 2).

High fidelityObtain up to 6-fold higher fidelity than Taq DNA Polymerase, ideal for cloning and microarray applications.

Increase product size and yieldAmplify very long PCR products from complex DNA templates with little or no optimization (Fig. 3). For PCR products greater than 2 kb, yields are greatly increased.

StableRepeated freeze-thaw cycles have no observed effect on performance (Figs. 1 and 3).

Functional tests:PCR with HotStart-IT FideliTaq Master Mix shifts production of primer-dimers to a specific target of 306 bp from 1 ng of human genomic DNA relative to FideliTaq Master Mix.

PCR with HotStart-IT FideliTaq Master Mix generates a 20.7 kb product.

HotStart-IT FideliTaq Master Mix Formulation (2X):HotStart-IT FideliTaq Master Mix combines FideliTaq DNA Polymerase with a recombinant hot start protein in a unique buffer formulation. Magnesium and nucleotide concentrations are 3 mM and 0.4 mM each, respectively.

Components:25 reactions 625 µl100 reactions 4 x 625 µl500 reactions 12.5 ml



91, 2216-2220.2. Cheng, S., Focklet, C., Barnes, W. M., and






HotStart-IT FideliTaq PCR Master Mix (2X)

Fig. 1. Increased specificity of the HotStart-IT FideliTaq PCR Master Mix and stability. A 306 bp fragment of the single-copy numb gene was amplified from 1 ng of human genomic DNA with and without HotStart-IT technology. The primers in this assay were designed with 3 bases of overlap at their 3'-ends to favor primer-dimer formation during reaction set-up at room temperature. Results demonstrate a shift from mainly primer-dimers to the desired product when HotStart-IT is used. Also, the mix is extremely stable as no loss in performance was observed following 10 freeze-thaw cycles (FTs) relative to a mix stored at -20°C.

Fig. 2. Sensitivity of the HotStart-IT FideliTaq PCR Master Mix. A 455 bp fragment of the single-copy numb gene was amplified from the indicated amounts of human genomic DNA. The master mix is extremely sensitive as amplification can be achieved from approxi-mately one human cell.

Fig. 3. Extreme amplicon length using HotStart-IT FideliTaq PCR Master Mix and stability. Two long PCR products of 20.7 kb and 35 kb were amplified from lambda DNA using 1 ng and 10 ng of DNA respectively. A 23 kb fragment of the β-globin gene was ampli-fied from 100 ng of human genomic DNA. A denaturation tempera-ture of 95°C was used for lambda targets and 92°C was used for the human target. Note that the mix is extremely stable as no loss in per-formance was observed following 10 freeze-thaw cycles (FTs) relative to a mix stored at -20°C.

500 bp –

100 bp –

no HotStart

HotStart

– numb, 306 bp

– primer-dimers

500 bp –

100 bp –

3 pg (1 cell)

30 pg

– numb, 455 bp

300 pg


PCR Tools | Hot Start PCR


1,000 units 5 x 250 units 5,000 unitsCustoms by request

Source:E. coli strain expressing a clone of Taq DNA Polym-erase from Thermus aquaticus(1-3). The hot start component is a recombinant protein also expressed in E. coli.

Description:HotStart-IT Taq DNA Polymerase uses a novel hot start method developed at Affymetrix called primer sequestration. In general, hot start PCR methods reduce or eliminate non-specific primer-extension products formed at lower temperatures during assembly of PCR reactions. At these less strin-gent annealing temperatures, primers may bind non-specifically, which often leads to unwanted amplification products and primer-dimers. In order to resolve this problem, we have combined Taq DNA Polymerase with a recombinant protein which binds and sequesters primers at lower tempera-tures making them unavailable for use by Taq DNA Polymerase. This primer-sequestration technique effectively blocks DNA synthesis from mis-priming events at lower temperatures (Fig. 1). Following the initial denaturation step during PCR, the protein is inactivated and the primers are free to participate in the amplification reaction. This novel hot start method enhances many complex PCR reactions by increasing both specificity and yield.

Applications:�� High-specificity PCR amplification�� High-sensitivity PCR amplification�� TA-vector cloning�� Amplification prior to in vitro transcription

Advantages:�� Room temperature reaction set-up�� High specificity and sensitivity�� Minimizes amplification of non-specific products and primer-dimers (Fig. 2)


Taq DNA Polymerase is a highly processive 5'→3' polymerase and has 5'→3' exonuclease activity. Suitable for TaqMan® assays.





Assay conditions:The reaction mixture (50 µl) contains 25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM 2-mercaptoethanol, 200 µM dNTPs, 250 µg/ml activated salmon sperm DNA, and cloned Taq DNA Polymerase. After incubation at 74°C for 10 minutes, acid-insoluble material is determined.

Polymerase blocking assay:The assay compares the polymerase activity of HotStart-IT Taq DNA Polymerase relative to Taq DNA Polymerase (PN 71160). The reaction mixtures contain 0.625 units of polymerase, 1X PCR Reaction Buffer, 0.2 mM each dNTP, and 2 pmol of overlap-ping, extendable oligonucleotides in a 25 µl reaction volume. Following incubation at 25°C for 4 hours, HotStart-IT Taq DNA Polymerase blocks at least 90% of the activity relative to Taq DNA Polymerase without hot start capability.

Functional test:PCR with HotStart-IT Taq DNA Polymerase shifts production of primer-dimers to a specific target of 306 bp from 1 ng of human genomic DNA relative to Taq DNA Polymerase.

Functionally tested 10X PCR Reaction Buffer (included):100 mM Tris-HCl (pH 8.6), 500 mM KCl, 15 mM MgCl2

Functionally tested MgCl2 (included):25 mM solution





3. Innis, M. A. and Gelfand, D. H. (1990) PCR Protocols: A Guide to Methods and Applications, Academic Press.

HotStart-IT Taq DNA Polymerase

100 pg DNA 1 ng DNA - - + + M - - + +

– numb, 306 bp– primer- dimers

Fig. 2. Increased specificity of HotStart-IT Taq DNA Polymerase. The single-copy numb gene was amplified from the indicated amounts of human genomic DNA with standard Taq DNA Polymerase (-) or with HotStart-IT Taq DNA Polymerase (+). Primers were designed with a 3 bp overlap at their 3'-ends to favor primer-dimer formation during reaction set-up at room temperature. Results demonstrate a shift from mainly primer-dimers to the desired product when HotStart-IT Taq DNA Polymerase is used.

Fig. 1. HotStart-IT Method: Primer Sequestration

PCR Reaction Preparation without Hot Start

• primers with self-complementary region

5 3

5 3

5 3

553

5 3

• primers hybridize to each other at low temperatures

PCR Reaction Preparation with USB Hot Start Method

5 3

5 3

53

5 3

5 3

53

• polymerases are recruited to hybrid• primer-dimers are produced• PCR Failure

• binding proteins interact with primers• primers are sequestered at low temperatures

• prevents non-specific hybridization and extension

• denaturation step inactivates binding proteins• primers bind to specific targets

• PCR Success

3Polymerase

Polymerase


Binding Proteins


71196 25 reactions100 reactions

500 reactionsCustoms by request

HotStart-IT Taq Master Mix uses a novel hot start method developed at Affymetrix called primer sequestration. In general, hot start PCR methods reduce or eliminate non-specific primer-extension products formed at lower temperatures during assembly of PCR reactions. At these less stringent annealing temperatures, primers may bind non-specifically, which often leads to unwanted amplification products and primer-dimers.

In order to resolve this problem, we have combined Taq DNA Polymerase with a recombinant protein which binds and sequesters primers at lower temperatures making them unavailable for extension by Taq DNA Polymerase. This primer-sequestration technique effectively blocks DNA synthesis from mis-priming events at lower temperatures (Fig. 1). Following the initial denaturation step during PCR, the protein is inactivated and the primers are free to participate in the amplification reaction. This novel hot start method enhances many complex PCR reac-tions by increasing both specificity and yield.

HotStart-IT Taq Master Mix is supplied as a conve-nient 2X pre-mixed formulation containing HotStart-IT Taq DNA Polymerase and Ultrapure nucleotides in a proprietary reaction buffer.

Applications:�� High-specificity PCR amplification�� High-sensitivity PCR amplification�� TA-vector cloning�� Amplification prior to in vitro transcription�� High-throughput PCR

ConvenientThe pre-mixed formulation saves time and reduces potential contamination errors by eliminating several pipetting steps. For a 50 µl reaction, simply add 25 µl of HotStart-IT Taq Master Mix to primers, DNA template, and PCR-Qualified H2O. Reactions can be easily performed in 10 µl, 25 µl, 50 µl or 100 µl volumes.

Room temperature reaction assembly is possible because of the hot start feature.

Novel hot start technologySince the polymerase is not chemically-inactivated, there is no extensive heating step necessary which reduces the chance of damaging precious DNA samples from heat-induced depurination.

Higher specificity and sensitivityHotStart-IT Taq Master Mix minimizes amplifica-tion of non-specific products and primer-dimers (Figs. 1 and 2). It amplifies PCR products with low background and from low-copy targets, essential for demanding genomic and cDNA applications with limited sample material.

StableThe mix withstands repeated freeze-thaw cycles and 4°C storage for extended periods of time with no observed decrease in performance (Fig. 2).

Functional test:PCR with HotStart-IT Taq Master Mix shifts produc-tion of primer-dimers to a specific target of 306 bp from 1 ng of human genomic DNA relative to Taq Master Mix.

HotStart-IT Taq Master Mix Formulation (2X):HotStart-IT Taq Master Mix combines Taq DNA Polymerase with a recombinant hot start protein in a unique buffer formulation. Magnesium and nucleotide concentrations are 3 mM and 0.4 mM each, respectively.

Components:25 reactions 625 µl100 reactions 4 x 625 µl500 reactions 12.5 ml



HotStart-IT Taq PCR Master Mix (2X)

– numb, 306 bp

– primer- dimers

M - + + + +-20

°CF/T 4°C RT

Fig. 2. Increased specificity and stability of HotStart-IT Taq Master Mix. Specificity: The single-copy numb gene was amplified from 1 ng of human genomic DNA with standard Taq Master Mix (-) or with HotStart-IT Taq Master Mix (+). Primers were designed with a 3 bp overlap at their 3'-ends to favor primer-dimer formation during reaction set-up at room temperature. Results demonstrate a shift from mainly primer-dimers to the desired product when HotStart-IT Taq Master Mix is used. Stability: The Master Mix shows no loss in performance following 15 freeze-thaw cycles (F/T), 4°C storage for one month (4°C), or room temperature storage for one month (RT) compared to a reference mix stored at -20°C.

4 hours at 24 hours at 25°C 25°C - Taq HS - Taq HS

– 50-mer extended product23-mer

template –

Fig. 1. HotStart-IT Taq Master Mix prevents primer exten-sion at room temperature. Two overlapping and extendable oligonucleotides were incubated in a mock PCR reaction at 25°C for either 4 or 24 hours with standard Taq Master Mix (Taq), HotStart-IT Taq Master Mix (HS), or no polymerase (-). Following incubation, reactions were separated on a 15% denaturing gel with urea. Results demonstrate that full-length extension of the HEX-labeled 23-mer to a 50-mer was completely blocked by HotStart-IT Taq Master Mix at room temperature while some extension occurred with the standard Taq Master Mix.

Related products


PCR Markers, 50 - 2000 bp76710 250 µl




71194 400 μgSufficient for 200 x 25 μl reactionsCustoms by request

Source:HotStart-IT Binding Protein is a recombinant protein expressed in E. coli.

Description:HotStart-IT Binding Protein is the active compo-nent in a novel hot start technology developed at Affymetrix called primer sequestration. In general, hot start PCR methods reduce or eliminate non-specific primer-extension products formed at lower temperatures during assembly of PCR reactions. At these less stringent annealing temperatures, primers may bind non-specifically, which often leads to unwanted amplification products and primer-dimers. In order to resolve this problem, we have produced a high-quality DNA binding protein that is especially useful at sequestering primers at lower temperatures making them unavailable for use by a polymerase (Fig. 1). This primer-sequestration technique effectively blocks DNA synthesis from mis-priming events at lower temperatures. Following the initial denaturation step during PCR, the protein is inactivated and the primers are free to participate in the amplification reaction. Unlike other hot start methods, such as antibodies or chemical modifica-tions, the binding protein is compatible with a vari-ety of thermostable polymerases. HotStart-IT Binding Protein has been designed for PCR applications that demand extremely high specificity and sensitivity and is thoroughly tested for purity and performance.

Application:�� Hot start PCR amplification

Properties:HotStart-IT Binding Protein performs well in many standard PCR reaction buffers.


Storage buffer:20 mM Tris-HCl (pH 8.5), 200 mM KCl, 0.1 mM EDTA, 1 mM DTT, and 50% glycerol.

Mass definition:In general, one microgram of HotStart-IT Binding Protein sequesters about 5 pmol of primers.

Concentration:≥ 10 mg/ml

Quality control polymerase blocking assay:This assay compares the amount of Taq DNA Polymerase activity with 2 μg HotStart-IT Binding Protein against its activity with no binding protein. The reaction mixtures contain 0.625 units of polymerase, 1X PCR Reaction Buffer (10 mM Tris- HCl [pH 8.6], 50 mM KCl, 1.5 mM MgCl2), 0.2 mM each dNTP, and 2 pmol of overlapping, extendable oligonucleotides in a 25 µl reaction volume. Following incubation at 25°C for 4 hours, HotStart-IT Binding Protein blocks at least 90% of the Taq DNA Polymerase activity.

Functional test:PCR with Taq DNA Polymerase and HotStart-IT Binding Protein shifts production of primer-dimers to a specific target of 306 bp from 1 ng of human genomic DNA relative to Taq DNA Polymerase alone.



HotStart-IT Binding Protein

Fig. 2. Increased specificity due to HotStart-IT Binding Protein. Single-copy numb and p53 genes were amplified from the indicated amounts of human genomic DNA using standard Taq DNA Polymerase without (-) and with (+) 2 µg of HotStart-IT Binding Protein per reaction. The primers in this assay were designed with 3 bases of overlap at their 3'-ends to favor primer-dimer formation dur-ing reaction set-up at room temperature. Results demonstrate a shift from mainly primer-dimers to the desired products when the binding protein was included.

Fig. 1. HotStart-IT Method: Primer Sequestration

PCR Reaction Preparation without Hot Start


5 3

5 3

5 3

553

5 3

• primers hybridize to each other at low temperatures

PCR Reaction Preparation with USB Hot Start Method

5 3

5 3

53

5 3

5 3

53

• polymerases are recruited to hybrid• primer-dimers are produced• PCR Failure

• binding proteins interact with primers• primers are sequestered at low temperatures

• prevents non-specific hybridization and extension

• denaturation step inactivates binding proteins• primers bind to specific targets

• PCR Success

3Polymerase

Polymerase


Binding Proteins

Advantages of HotStart-IT:�� Room temperature reaction set-up�� High specificity and sensitivity�� Minimizes amplification of non-specific products and primer-dimers (Fig. 2).�� Ideal for complex templates and multiplex reactions�� Unlike chemically-modified Taq, no extensive heating step is necessary which may damage precious samples.�� Technology is portable to a polymerase of choice.

– numb, 306 bp– primer-dimers

– p53, 1142 bp

– primer-dimers

- - + + M - - + +

- - + + M - - + +


PCR Tools | Multiplex PCR


MagniTaq Multiplex PCR Master Mix is a single-tube, 2X multiplex PCR master mix with validated perfor-mance for simultaneous amplification of at least 20 targets up to 1 kb in length of difficult templates with minimal optimization. MagniTaq Multiplex PCR Master Mix (2X) contains hot start MagniTaq DNA Polymerase, dNTPs, and MgCl2 in a proprietary buffer formulation designed to enable multiplex PCR without lengthy optimization procedures.

Features:�� Advanced buffer formulation that includes dNTPs and MgCl2, ensuring efficient, rapid, multiplex PCR without lengthy optimization procedures�� Validated performance for simultaneous amplification of at least 20 targets up to 1 kb in length�� Quality results on difficult sample types; superior data even from FFPE samples or for single-copy genes�� Single-tube, pre-mixed solution. Simply add template, primers, and water

Applications:�� End point PCR of multiple targets simultaneously�� Minimizes effect of off-target priming events

Reliable and efficient multiplexing of over 20 plexesIn order to enhance the specificity and sensitivity of multiplexing, the mix uses MagniTaq DNA Polymerase, a chemically modified, hot start polymerase that is completely inactive at ambient temperatures. MagniTaq DNA Polymerase minimizes the effect of off-target priming events, such as primer-dimers, which often occur at lower tempera-tures during reaction set-up and prior to the initial denaturation step.

In addition, MagniTaq Multiplex PCR Master Mix efficiently amplifies greater than 20 products simul-taneously over a broad range of template amounts and primer concentrations.

Data generated shows 22-plex amplification from single-copy genes in the Notch signaling pathway. Even at template amounts ranging from 1–100 ng of human gDNA, amplification of 22 targets is observed (Fig. 1). The reaction was done without optimization in a single tube of pre-mixed solution of a proprietary buffer that includes dNTPs and MgCl2.

Varying primer concentrations can affect the ef-ficiency and amplification effectiveness. Typically, lower primer concentrations favor longer products, while higher primer concentrations favor shorter products. MagniTaq Multiplex PCR Master Mix is designed to work over a range of primer concentra-tions (Fig. 2).

Primer design�� The length of primers should be 20-30 bases.�� Aim for a Tm equal to or greater than 60°C; primer sets should be designed to have similar melting temperatures if possible.�� Optimal GC content should be 40-65%.�� Avoid designing primers with 3 or more G or C residues sequentially near the 3’ end, as well as primers that can form hairpins or that have significant complementary sequence to one another.�� Typically, lower primer amounts tend to favor longer targets, while higher primer amounts tend to favor shorter targets.

MagniTaq™ Multiplex PCR Master Mix (2X)

Fig. 1. 22-plex amplification of low amounts of gDNA. 22-plex amplification of gDNA targets with MagniTaq Multiplex PCR Master Mix. 1, 10, 100, and 1000 ng human genomic DNA input amounts (1 ng is about 100 cells) are added to the 22-plex notch-pathway single-copy gene primer set. Primer concentration is 0.2 µM and samples were loaded with SYBR® Green dye and run on an agarose gel.

1 10 100 1000 ng template DNA

1000900800700600

500

400

300

200

100

1252 1252

142 142

1500

1000

750

500

300

150

50 Fig. 2. 22-plex of a wide range of primer concentrations. MagniTaq Multiplex PCR Master Mix is effective over a wide range of primer concentrations. 100 ng human genomic DNA input amount is added to the 22-plex notch-pathway, single-copy gene primer set. Duplicate reactions were run using 0.1, 0.2 or 0.4 µM primer concentrations. Samples were loaded with SYBR Green dye and run on an agarose gel.

µM primers: 0.1 0.2 0.4

1000900800700600

500

400

300

200

100

1252

142

1500

1000

750

500

300

150

50



Fig. 4. Comparison testing of 22-plex FFPE samples. 22-plex FFPE sample amplification competitor comparison data of normal vs tumor sample types. 100 ng human genomic DNA input from 2008 breast FFPE samples and 2002 lung FFPE samples, normal or tumor, are added to the 22-plex notch pathway primer set. 0.1 µM primer sets are used. Comparison of MagniTaq Multiplex PCR Master Mix (MT) to competitor Q and QP were run. Samples are loaded with SYBR Green and run on 2% TAE-agarose gel.

Fig. 3. Superior multiplexing of FFPE samples. 20-plex FFPE sample amplification competitor comparison data of normal vs. tumor sample types. 25 ng human genomic DNA input from 2008 breast FFPE samples and 2002 lung FFPE samples, normal or tumor, are added to the 20-plex notch pathway primer set. 0.2 µM primer sets are used. Comparison of MagniTaq Multiplex PCR Master Mix (MT) to competitor Q and QP were run. Samples are loaded with SYBR Green and run on 2.25% TAE-agarose gel.

Breast—2,008 Lung—2,002

50 ng FFPE gDNANormal (N) or Tumor (T)

MT Q QP MT Q QP N T N T N T N T N T N T

1000900800700600500400300

200

100

1500

1000

750

500

300

150

50

1252

142

25 ng FFPE gDNANormal (N) or Tumor (T)

MT Q QP MT Q QP N T N T N T N T N T N T

MagniTaq™ Multiplex PCR Master Mix (2X) continued...

Superior performance on difficult targets with minimal optimizationMagniTaq Multiplex PCR Master Mix offers quality results on difficult to amplify sample types. Genomic DNA isolated from FFPE tissue samples is often degraded, and therefore, difficult to amplify using standard PCR reagents. Multiplex PCR with these sample types calls for an exceptional amount of optimization to produce consistent amplification of all sought after targets.

The uniform product yield obtained when using MagniTaq Multiplex PCR Master Mix allows for more products to be amplified, especially when compared to mixes from other suppliers (Figs. 3 and 4). The mixes from other suppliers show a much more pronounced decrease in the amplification of the high molecular weight products compared to MagniTaq Multiplex PCR Master Mix (Figs. 3 and 4). MagniTaq Multiplex PCR Master Mix is, therefore, more suit-able for PCR amplification of multiple primer sets from degraded FFPE gDNA template, particularly for older tissue samples.

Components:25 reactions—for 25 x 50 μl multiplex PCR

reactions100 reactions—for 100 x 50 μl multiplex PCR

reactions

Shipping and storage:Shipped on dry ice. For long-term storage, store at -20°C. When using, the mix may be stored at 4°C for up to 2 months.



MP1100 kitPCR Primers*MP11XX *Use digits from the PCR Primers product number. See chart below for full listing of PCR Primers.

�� Quickly and easily detect the methylation status of your promoter of interest�� No hassle of sodium bisulfate modification�� Choose from our list of 40 PCR primers OR use your own PCR primers to detect methylation of promoters specific for your research.

Use the Promoter Methylation PCR Kit to quickly and easily detect the methylation status of your promoter of interest, without the hassle of chemical modifica-tion with sodium bisulfate.

Standard molecular biology techniques do not preserve methylation of the genomic DNA, causing a hindrance in the appreciation of methylation as an important epigenetic event in cancer progression. Affymetrix has developed a PCR method for analyz-ing methylation status of a specific promoter of interest. The method is based on MeCP2 bind-ing, which differentiates those promoters with methylated groups from unmethylated promoters. Compared to sodium bisulphate-based conversion of methylated bases in conjunction with PCR amplifica-tion, this method is very simple and straightforward.

The flowchart below details how this procedure works:Isolated genomic DNA is digested with Mse I, and the resulting DNA fragments are incubated with the

methylation binding protein MeCP2 (a.k.a. MBP). The methylated DNA fragments are isolated with a spin column and then amplified with promoter specific primers. Agarose gel electrophoresis is used to visualize the PCR products. The presence of a band on the gel indicates that a specific promoter is methylated in your genomic DNA sample.

Mse I and PCR enzymes are not included in this kit. We recommend HotStart-IT FideliTaq DNA Polymerase, PN 71155 and MagniTaq Multiplex PCR Master Mix, PN 71199, for PCR (see below).

Promoter Methylation PCR Kit

PCR primers:Gene Name Forward Reverse Approx. PCR Prod. No. Primer Position Primer Position Product Length14-3-3 1 180 bp MP1101 “ 180 “ MP1102

14-3-3 300 600 bp MP1103 “ 900 “ MP1104

ABL1 2 390 bp MP1105 “ 400 “ MP1106

BAGE 1 230 bp MP1107 “ 230 “ MP1108

BAGE 260 490 bp MP1109 “ 650 “ MP1110

BAGE 670 230 bp MP1111 “ 900 “ MP1112

BRCA1 50 200 bp MP1113 “ 250 “ MP1114

BRCA1 550 130 bp MP1115 “ 680 “ MP1116

Calcitonin CGPR 1 300 bp MP1117 “ 450 “ MP1118

Calcitonin CGPR 670 200 bp MP1119 “ 900 “ MP1120

CD14 540 400 bp MP1121 “ 1000 “ MP1122

CDKN2A 900 200 bp MP1123 “ 1100 “ MP1124

Gene Name Forward Reverse Approx. PCR Prod. No.Primer Position Primer Position Product Length

CDKN2A 270 240 bp MP1125 “ 510 “ MP1126

COX2 280 320 bp MP1127 “ 601 “ MP1128

CyclinD2 580 170 bp MP1129 “ 750 “ MP1130

CyclinD2 440 130 bp MP1131 “ 570 “ MP1132

DAPK 50 200 bp MP1133 “ 250 “ MP1134

DAPK 400 500 bp MP1135 “ 900 “ MP1136

glut4 190 200 bp MP1137 “ 390 “ MP1138

H-ras 40 400 bp MP1139 “ 460 “ MP1140

H-ras 500 300 bp MP1141 “ 840 “ MP1142

INFg 500 100 bp MP1143“ 600 “ MP1144

GAGE1 100 250 bp MP1145 “ 390 “ MP1146

GAGE1 400 400 bp MP1147 “ 800 “ MP1148

Fig. 1. Methylation PCR verification of array results. Results obtained with the Promoter Methylation Array can be verified with the Promoter Methylation PCR Kit. Genomic DNA from MCF7 and HeLa cells was analyzed with the Promoter Methylation Array (top). The promoters (indicated as boxes) were verified by individual meth-ylation PCR. Left band: MCF7. Right band: HeLa. (bottom).

Related products

HotStart-IT FideliTaq DNA Polymerase71155 50 units

250 units1,000 units5,000 units

MagniTaq Multiplex PCR Master Mix (2X)71199 25 reactions

100 reactions



Promoter Methylation PCR Kit continued...



Source:All proteins are recombinant versions expressed in E. coli.

Description:HotStart-IT FideliTaq DNA Polymerase combines a novel hot start method developed at Affymetrix with the long and accurate amplification properties of FideliTaq DNA Polymerase. The new hot start method, called primer sequestration, uses a binding protein to reduce or eliminate non-specific primer-extension products which may be generated at lower temperatures during assembly of PCR reactions. Following the initial denaturation step during PCR, the binding protein is inactivated and the primers are free to participate in the amplification reaction. This novel method enhances many complex PCR reactions by increasing specificity and yield, as well as sensitivity.

FideliTaq DNA Polymerase combines Taq DNA Polym-erase with a high-fidelity, proofreading polymerase. This enzyme blend has the 5'→3' exonuclease activity of Taq DNA Polymerase as well as the 3'→5' exonuclease activity of the proofreading enzyme. FideliTaq DNA Polymerase increases fidelity up to 6-fold over Taq DNA Polymerase alone and allows for amplification of longer products(1-4). The enzyme blend generates PCR products whose ends are compatible with either blunt-end or TA cloning procedures(5), with A-tailed ends favored over blunt ends in an approximately 3 to 1 ratio.

FideliTaq DNA Polymerase can be used in conjunc-tion with the Promoter Methylation PCR Kit for gene amplification prior to methylation detection.

Applications:�� High-specificity and high-sensitivity PCR amplification�� Extremely long PCR amplification�� PCR-mediated cloning

Advantages:�� Room temperature reaction set-up.�� Minimizes amplification of non-specific products and primer-dimers.�� High specificity and sensitivity �� Generates long PCR amplicons�� Up to 6-fold higher fidelity than Taq polymerase.



Storage buffer:20 mM Tris-HCl (pH 8.5), 1 mM DTT, 0.1 mM EDTA, 200 mM KCI, 50% glycerol, and stabilizers.



Assay conditions:The reaction mixture contains 25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM 2-mercaptoethanol, 200 µM dNTPs, 250 µg/ml activated salmon sperm DNA, and Taq DNA Polymerase. Following incubation at 74°C for 10 minutes, acid-insoluble material is determined.

Functional tests:PCR with HotStart-IT FideliTaq DNA Polymerase shifts production of primer-dimers to a specific target of 306 bp from 1 ng of human genomic DNA rela-tive to FideliTaq DNA Polymerase.

PCR with HotStart-IT FideliTaq DNA Polymerase generates a 20.7 kb product.

Functionally tested 10X PCR Reaction Buffer (included): 100 mM Tris-HCl (pH 8.6), 500 mM KCl, 15 mM MgCl2Functionally tested MgCl2 (included):25 mM solution



91, 2216-2220. 2. Cheng, S., Fockler, C., Barnes, W. M., and


3. Barnes, W. M. (1992) Gene 112, 29-35. 4. Cline, J., Braman, J. C., and Hogrefe, H. H. (1996)

Nucl. Acids Res. 24, 3546-3551. 5. Magnuson, V. L., Ally, D. S., Nylund, S. J.,




HotStart-IT FideliTaq DNA Polymerase

PCR primers continued:Gene Name Forward Reverse Approx. PCR Prod. No. Primer Position Primer Position Product LengthHOXA2 1 300 bp MP1149 “ 300 “ MP1150

IL4 450 230 bp MP1151 “ 680 “ MP1152

IRF7 350 450 bp MP1153 “ 850 “ MP1154

NIS 1 350 bp MP1155 “ 400 “ MP1156

NIS 500 300 bp MP1157 “ 800 “ MP1158

NME2 80 170 bp MP1159 “ 250 “ MP1160

NME2 280 500 bp MP1161 “ 800 “ MP1162

PDGFB 1 500 bp MP1163 “ 678 “ MP1164

Gene Name Forward Reverse Approx. PCR Prod. No.Primer Position Primer Position Product Length

POMC 50 490 bp MP1165 “ 540 “ MP1166

NES 150 290 bp MP1167 “ 410 “ MP1168

NES 450 350 bp MP1169 “ 800 “ MP1170

PRA2 400 250 bp MP1171 “ 650 “ MP1172

TFF1 510 160 bp MP1173 “ 750 “ MP1174

VHL 80 290 bp MP1175 “ 380 “ MP1176

Survivin 1 300 bp MP1177 “ 300 “ MP1178

CASP8 180 360 bp MP1179 “ 640 “ MP1180


PCR Tools | Real-Time qPCR


1,000 units 5,000 units

�� Chemically-modified Taq polymerase for hot start PCR�� Minimizes the amplification of non-specific products and primer dimers.�� Exceptional performance, offering high specificity and sensitivity�� Room temperature reaction set-up

VeriQuest Taq DNA Polymerase is a highly purified, hot start Taq polymerase that has no polymerase activity prior to the initial heat activation step, ensur-ing there is no amplification of non-specific primers, such as primer-dimers. This feature allows the con-venience of reaction assembly at room temperature as well as higher specificity and sensitivity. The initial incubation step of 95°C for 10 minutes before PCR cycling removes the blocking chemical moiety, result-ing in activation of the polymerase.

Properties:VeriQuest Taq DNA Polymerase is a chemically-modified, full length Taq DNA polymerase for hot start.


Storage buffer:20 mM Tris-HCl, pH 8.5, 100 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% glycerol and stabilizers.

Unit definition: One unit of enzyme is defined as the amount that catalyzes the incorporation of 10 nmol of total nucleotide into acid-insoluble form in 30 minutes at 74°C.

Assay conditions: The reaction mixture contains 25 mM TAPS, pH 9.3 (at 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM β-ME, 200 µM each dATP, dGTP, dTTP, 100 µM [α-32P]-dCTP (0.05 to 0.1 Ci/mmole), 250 µg/ml activated salmon

sperm DNA, and VeriQuest Taq DNA Polymerase. After incubation at 74°C for 10 minutes, acid insoluble material is determined (50 µl reaction volume).

Functional test: Functionally tested for PCR according to the stan-dard Affymetrix PCR protocol. This protocol is based upon the amplification of a 500 bp lambda DNA.

Concentration: 5 units/µl

Functionally tested VeriQuest Taq 10X PCR Reaction Buffer (included):150 mM Tris-HCl, pH 8.0, 500 mM KCl, 15 mM MgCl2Functionally tested MgCl2 (included):25 mM MgCl2 solution

Storage:Shipped on dry ice. Store at -20°C.

VeriQuest Taq DNA Polymerase

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SYBR Green mixes

VeriQuest SYBR Green qPCR Master Mix (2X)75600 40 reactions

200 reactions 400 reactions 1,000 reactions 2,000 reactions

VeriQuest SYBR Green qPCR Master Mix with Fluorescein (2X)75665 40 reactions


Probe mixes

VeriQuest Probe qPCR Master Mix (2X)75650 40 reactions


VeriQuest Probe qPCR Master Mix, No Reference Dye (2X)75660 40 reactions


Fast SYBR Green mixes

VeriQuest Fast SYBR Green qPCR Master Mix (2X)75690 100 reactions 500 reactions 1,000 reactions 2,500 reactions 5,000 reactions

VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein (2X)75675 100 reactions 500 reactions 1,000 reactions 2,500 reactions 5,000 reactions

Fast Probe mixes

VeriQuest Fast Probe qPCR Master Mix (2X)75680 100 reactions 500 reactions 1,000 reactions 2,500 reactions 5,000 reactions

VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X)75685 100 reactions 500 reactions 1,000 reactions 2,500 reactions 5,000 reactions

One-Step qRT-PCR SYBR Green mixes

VeriQuest SYBR Green One-Step qRT-PCR Master Mix (2X)75705 40 reactions 200 reactions

VeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein (2X)75715 40 reactions 200 reactions

One-Step qRT-PCR Probe mixes

VeriQuest Probe One-Step qRT-PCR Master Mix (2X)75700 40 reactions 200 reactions

VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye (2X)75710 40 reactions 200 reactions


75600 (50 µl reaction volume) 40 reactions (1 ml) 200 reactions (5 ml) 400 reactions (2 x 5 ml) 1,000 reactions (5 x 5 ml) 2,000 reactions (10 x 5 ml)

VeriQuest SYBR Green qPCR Master Mix is supplied as a 2X pre-mixed formulation containing both ROX and UDG in an optimized buffer for quality results in real-time quantitative PCR assays. The proprietary re-action buffer and the hot start polymerase enhance SYBR Green-based qPCR amplification by reducing primer-dimer formation and non-template priming, increasing specificity and sensitivity. The master mix contains a chemically-modified Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), SYBR Green I, and ROX Passive Reference Dye in a proprietary reaction buffer.

Features:�� One-tube master mix with ROX Reference Dye included�� Same protocols and same primers for standard mode amplification�� Consistency over a broad dynamic range: 7 orders of magnitude linear detection range with a correlation coefficient ≅ 1 (see Fig. 1) �� Exceptional performance with challenging GC-rich regions (see Fig. 3)�� Detect less than 4 copies of target from genomic DNA and differentiate less than a two-fold difference.�� Contains dUTP and Uracil-DNA Glycosylase (UDG or UNG) for carryover contamination prevention.�� Hot start PCR for room temperature reaction setup�� Stable at room temperature for 72 hours in a preassembled reaction

Reproducible, consistent results while main-taining precision and efficiencyThe convenience of a master mix formulation without sacrificing quality. VeriQuest SYBR Green qPCR Master Mix provides consistency over a broad dynamic template concentration range allowing 7 orders of magnitude linear detection (Fig. 1). High precision in target quantification and discrimination of a 1.33 to 10-fold dilution series with VeriQuest SYBR Green qPCR Master Mix ensure accurate results.

Enhanced amplification sensitivity and accuracy of low copy number detectionReliable detection from as little as 4 copies of a single copy gene and differentiation from less than a two-fold difference (Fig. 2).

Exceptional performance with challenging templatesEven in high GC and AT rich regions, VeriQuest SYBR Green qPCR Master Mix offers exceptional performance and high specificity so you have the confidence to verify your gene expression results (Figs. 3 and 4).

Ready-to-use, single tube replacementThe one tube master mix contains all necessary com-ponents: chemically-modified Taq DNA polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), SYBR Green I, and ROX as the passive reference dye in a proprietary reaction buffer. Simply add your DNA template, primers, and water, and you can begin your qPCR reaction. For use in your new or existing protocols, on any leading PCR platform, providing easy transition from other master mixes.

Passive Reference Dye and UDG includedROX Passive Reference Dye is an inert dye, whose fluorescence does not change during the reaction. This signal normalization is necessary to correct for well-to-well differences that may occur due to artifacts such as pipetting errors or sample evapora-tion. Uracil-DNA Glycosylase (UDG or UNG) and dUTP offer an option for carry-over contamination prevention from previous PCR amplifications. All are at optimized levels so there is no need to adjust concentrations.

Highly stable and easy to work with VeriQuest SYBR Green qPCR Master Mix is stable at room temperature for 72 hours in a pre-assembled reaction and can be stored at 4°C for convenient handling (Figs. 5 and 6); no lost time waiting for your master mix to thaw. Testing of 10 freeze-thaw cycles showed no loss in master mix performance. Ideal for high-throughput handling.

Components:40 reactions: 1 ml200 reactions: 5 ml400 reactions: 2 x 5 ml1,000 reactions: 5 x 5 ml2,000 reactions: 10 x 5 ml

References:1. VanGuilder, H. D., Vrana, K. E., and Freeman, W. M.

(2008) BioTechniques 44 (5), 619-626.2. Longo, M. C., Berninger, M. S., and Hartley, J. L.

(1990) Gene 93, 125-128.3. Tewhey, R. et al. (2009) Nature Biotechnology 27,

1025-1031.

VeriQuest SYBR Green qPCR Master Mix (2X)


Fig. 2. High precision in target quantification with VeriQuest SYBR Green qPCR Master Mix. Amplification plot from real-time PCR for a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNAs reverse-transcribed from HeLa total RNA.

Fig. 1. Linear detection range of VeriQuest SYBR Green qPCR Master Mix. Real-time amplification plot from a 10-fold dilu-tion series of a GAPDH synthetic target with starting amounts of 109 copies amplified in four replicate reactions using the ABI StepOne™ Real-Time PCR System.

(Continued on next page)



VeriQuest SYBR Green qPCR Master Mix continued...

Fig. 4. Melt curve from a 71.2% GC target amplicon. Human male genomic DNA was used.

Fig. 3. Amplification of GC-rich target with VeriQuest SYBR Green qPCR Master Mix. Standard curves from a 10-fold dilution series of 100 ng to 10 pg human male genomic DNA amplified in four replicate reactions with VeriQuest SYBR Green qPCR Master Mix, BioRad iTaq™ Fast SYBR Green Supermix with ROX and ABI Power® SYBR Green PCR Master Mix using the ABI 7500 PCR System for detection of a 71.2% GC-target amplicon (PROC, NT_022135.16)(3).

USB VeriQuest

BioRad

ABI

-2.5 -1.5 -0. 5 0.5 1.5 2.5

log gDNA input

44

39

29

34

Ct

19

24

PROC, 71.2% GCMix USB VQ BioRad ABIR2 0.999 0.944 0.997Slope -3.36 -1.75 -3.28EPCR 98.2% 272.6% 101.7%

Fig. 5. VeriQuest SYBR Green qPCR Master Mix stability for reliable performance of high-throughput handling. Pre-assembled PCR reaction stability after 48 and 72 hour incubations at room temperature from quantification of GADPH cDNAs.

Fig. 6. Inset defining Ct levels of 1 ng and 10 pg of cDNAs reverse transcribed from HeLa total RNA. GADPH was detect-ed from preassembled PCR reactions incubated at room temperature for 48 and 72 hours.

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200 reactions400 reactions

1,000 reactions2,000 reactions

VeriQuest Fast SYBR Green qPCR Master Mix (2X)75690 100 reactions

500 reactions1,000 reactions2,500 reactions5,000 reactions

VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein (2X)75675 100 reactions


First-Strand cDNA Synthesis Kit for Real-Time PCR75780 50 reactions

VeriQuest Taq DNA Polymerase71170 50 units


Water, Nuclease-Free 71786 10 x 1 ml

100 ml





VeriQuest SYBR Green qPCR Master Mix with Fluo-rescein is supplied as a 2X pre-mixed formulation containing both fluorescein and UDG in an opti-mized buffer for quality results in real-time quantita-tive PCR assays. The proprietary reaction buffer and the hot start polymerase enhance SYBR Green-based qPCR amplification by reducing primer-dimer forma-tion and non-template priming, increasing specificity and sensitivity.

Features:�� One-tube master mix with fluorescein added in optimized levels for use in all thermo cyclers that utilize it for signal normalization�� Enhanced amplification sensitivity and accuracy of low copy number detection�� Reproducible, consistent results while maintaining precision and efficiency�� Exceptional performance with challenging templates�� Ready-to-use, single-tube replacement with UDG included�� Highly stable and easy to work with

Enhanced sensitivity on amplification for low copy number detectionReliable detection from as little as four copies of a single copy gene and differentiation from less than a two-fold difference.

Reproducible, consistent results while main-taining precision and efficiencyThe convenience of a master mix formulation with-out sacrificing quality. VeriQuest SYBR Green qPCR Master Mix with Fluorescein provides consistency over a broad dynamic template concentration range allowing 7 orders of magnitude linear detec-tion. High precision in target quantification and discrimination of a 1.33 to 10-fold dilution series with VeriQuest SYBR Green qPCR Master Mix with Fluorescein ensures accurate results.

Exceptional performance with challenging templatesEven in high GC and AT rich regions, VeriQuest SYBR Green qPCR Master Mix with Fluorescein offers excep-tional performance and high specificity so you have the confidence to verify your gene expression results.

Ready-to-use, single-tube replacementThe one-tube master mix contains all necessary components: chemically-modified Taq DNA polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), SYBR Green I, and Fluorescein, as the passive reference dye, in a proprietary reaction buffer. Simply add your DNA template, primers, and water and you can begin your qPCR reaction. For use with instruments requiring fluorescein as a passive reference dye, i.e., Bio-Rad iCycler iQ®/ iQ5, Bio-Rad iCycler MyiQ™.

Passive reference dye and UDG includedFluorescein Passive Reference Dye is an inert dye whose fluorescence does not change during the reaction. This signal normalization is necessary to correct for well-to-well differences that may occur due to artifacts such as pipetting errors or sample evaporation.

Uracil-DNA Glycosylase (UDG or UNG) and dUTP offer an option for carry-over contamination prevention from previous PCR amplifications. All are at optimized levels so there is no need to adjust concentrations.

Highly stable and easy to work withVeriQuest SYBR Green qPCR Master Mix with Fluorescein is stable at room temperature for 72 hours in a preassembled reaction. This mix can be stored at 4°C for convenient handling, with no time lost waiting for your master mix to thaw. Testing of 10 freeze-thaw cycles showed no loss in master mix performance. This master mix is ideal for high-throughput handling.

Components:40 reactions: 1 ml200 reactions: 5 ml400 reactions: 2 x 5 ml1,000 reactions: 5 x 5 ml2,000 reactions: 10 x 5 ml

Storage:Shipped on dry ice. Store at -20°C for long-term storage. Store at 4-8°C for short-term storage (≤3 months).

VeriQuest SYBR Green qPCR Master Mix with Fluorescein (2X)


Amplification plots and standard curves from real-time PCR for dilution series of a synthetic GAPDH target with starting amounts of 108 copies amplified in triplicate reactions using the Bio-Rad MyiQ™ Single Color Real-Time PCR Detection System.

Linear detection range of VeriQuest SYBR Green qPCR Master Mix with Fluorescein


75690 (20 µl reaction volume)100 reactions (1 ml)500 reactions (5 ml)

1,000 reactions (2 x 5 ml)2,500 reactions (5 x 5 ml)

5,000 reactions (10 x 5 ml)

Features:�� Uses fast mode thermal cycling conditions for results in one-third of the time when compared to standard run protocols.�� One-tube master mix with ROX Reference Dye included – just add template, primers, and water �� Exceptional performance with challenging GC-rich regions�� Contains dUTP and Uracil-DNA Glycosylase (UDG or UNG) for carryover contamination prevention.�� Hot start PCR for room temperature reaction setup�� Sensitivity and precision with limited targets

VeriQuest Fast SYBR Green qPCR Master Mix is op-timized for SYBR Green detection on all instruments that utilize ROX as a passive reference dye and can be run using Fast mode cycling protocols. The 2X ready-to-use master mix contains hot start VeriQuest Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), SYBR Green, and ROX Passive Reference Dye in a proprietary reaction buf-fer. The hot start Taq polymerase has no polymerase activity prior to the initial heat activation step, which allows reaction assembly at room temperature, as well as higher specificity and sensitivity. Since the mix contains dUTP and UDG, carryover contamination

prevention can be performed prior to amplification. VeriQuest Fast SYBR Green qPCR Master Mix offers the same sensitivity and specificity found with our standard mode mixes, producing results in a fraction of the time.

VeriQuest Fast SYBR Green qPCR Master Mix shows exceptional efficiency and high specificity on challenging templates such as high GC- and AT-rich regions. The optimized formulation ensures no sacrifice in quality for increased speed. Performance comparisons highlight the sensitivity and reliable performance of this platform-independent solution. The sensitivity of the master mix allows for discrimi-nation from a 1.33-fold difference in gene target amount detected. In Fig. 2, a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNA reverse-transcribed from HeLa total RNA were amplified with an ef-ficiency of >99%.

VeriQuest Fast SYBR Green qPCR Master Mix is highly stable and easy to work with. It remains stable at room temperature for 72 hours in a pre-assembled reaction and can be stored at 4°C for convenient handling. The mix also allows for room temperature reaction set-up. The speed and stability make VeriQuest Fast SYBR Green qPCR Master Mix ideal for high-throughput handling.

Components:100 reactions 1 ml500 reactions 5 ml1,000 reactions 2 x 5 ml2,500 reactions 5 x 5 ml5,000 reactions 10 x 5 ml

Storage conditionsStore at -20°C for long-term storage. Store at 4-8°C for short-term storage (≤ 3 months).


VeriQuest Fast SYBR Green qPCR Master Mix (2X)

Fig.1. Linear detection range of VeriQuest Fast SYBR Green qPCR Master Mix. Amplification plot and standard curve from real-time PCR for a dilution series of a synthetic target with starting amounts of 109 copies amplified in four replicate reactions using the ABI 7500 Real-Time PCR System and GAPDH primers in fast mode with 5 minutes activation at 95°C. The amplification process was lin-ear over eight orders of magnitude.

Fig. 2. High sensitivity and precision in limited target quantification. Amplification plot (left) and standard curve (right) from real-time PCR for a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNAs reverse-transcribed from HeLa total RNA.

GAPDH Kras

10 43 2 1 ng RNA

10 4 3 2 1 ng RNA

GAPDH2y = -3.3773x + 22.224

EPCR = 97.7%R² = 0.9955

Krasy = -3.2461x + 30.591

EPCR = 103.3%R² = 0.9993

151719212325272931

-0.2 0 0.2 0.4 0.6 0.8 1 1.2

Ct

log input

GAPDH2 Kras


15

20

25

30

35

-4.0 -2.0 0.0 2.0

Ct

log ng input

B2M

ABI Fast SYBR

Bio-Rad iTaq Fast

VeriQuest Fast SYBR

VeriQuest Fast SYBRy = -3.1802 + 23.205EPCR = 106.3%R2 = 0.9999

Bio-Rad iTaq Fast SYBRy = -3.3764 + 22.908EPCR = 97.8%R2 = 0.9991

ABI Fast SYBRy = -3.1984 + 23.755EPCR = 105.4%R2 = 0.9995

Fig. 3. Exceptional results with performance comparison. Standard curves from real-time PCR for a 10-fold dilution series of 10 ng to 1 pg cDNA reverse-transcribed from HeLa total RNA amplified in duplicate reactions with VeriQuest Fast SYBR Green qPCR Master Mix, Bio-Rad iTaq Fast SYBR Green Supermix with ROX™, and ABI Fast SYBR Green Master Mix.






VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein (2X)

75675 (20 µl reaction volume)100 reactions (1 ml)

500 reactions (5 ml) 1,000 reactions (2 x 5 ml) 2,500 reactions (5 x 5 ml) 5,000 reactions (10 x 5 ml)

Features:�� Uses fast mode thermal cycling conditions for results in one-third of the time when compared to standard run protocols.�� One-tube master mix with fluorescein passive reference dye included – for use in BioRad thermocyclers �� Exceptional performance with challenging GC-rich regions�� Contains dUTP and Uracil-DNA Glycosylase (UDG or UNG) for carryover contamination prevention.�� Hot start PCR for room temperature reaction setup�� Sensitivity and precision with limited targets

VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein is optimized for SYBR Green detec-tion on BioRad instruments that utilize fluorescein as a passive reference dye and can be run using Fast mode cycling protocols. The 2X ready-to-use master mix contains hot start VeriQuest Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), SYBR Green, and Fluorescein Passive Reference Dye in a proprietary reaction buffer. The hot start Taq polymerase has no polymerase activity prior to the initial heat activation step, which allows reaction assembly at room temperature, as well as higher specificity and sensitivity. Since the mix contains dUTP and UDG, carryover contamination prevention can be performed prior to amplification. VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein offers the same sensitivity and specificity found with our standard mode mixes producing results in a fraction of the time.

VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein shows exceptional efficiency and high specificity on challenging templates such as high GC- and AT-rich regions. The optimized formulation ensures no sacrifice in quality for increased speed. Performance comparisons highlight the sensitiv-ity and reliable performance of this solution.The sensitivity of the master mix allows for discrimination from a 1.33-fold difference in gene target amount detected. In Fig. 2, a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNA reverse-transcribed from HeLa total RNA was amplified with an efficiency of >99%.

VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein is highly stable and easy to work with. It remains stable at room temperature for 72 hours in

a pre-assembled reaction and can be stored at 4°C for convenient handling. The mix also allows for room temperature reaction set-up. The speed and stability make VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein ideal for high-through-put handling.


Storage conditionsStore at -20°C for long-term storage. Store at 4-8°C for short-term storage (≤3 months).


Fig. 2. High sensitivity and precision in limited target quantification. Amplification plot (left) and standard curve (right) from real-time PCR for a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNAs reverse-transcribed from HeLa total RNA.

GAPDH Kras

10 43 2 1 ng RNA

10 4 3 2 1 ng RNA

GAPDH2y = -3.3773x + 22.224

EPCR = 97.7%R² = 0.9955

Krasy = -3.2461x + 30.591

EPCR = 103.3%R² = 0.9993

151719212325272931

-0.2 0 0.2 0.4 0.6 0.8 1 1.2

Ct

log input

GAPDH2 Kras


15

20

25

30

35

-4.0 -2.0 0.0 2.0

Ct

log ng input

B2M

ABI Fast SYBR

Bio-Rad iTaq Fast

VeriQuest Fast SYBR











HotStart-IT SYBR Green qPCR Master Mix uses a novel hot start method developed at Affymetrix called primer sequestration. With this method, an oligo binding protein sequesters primers at lower temperatures making them unavailable for use by Taq DNA Polymerase. Following the initial denatur-ation step, the binding protein is inactivated and the primers are released. HotStart-IT SYBR Green qPCR Master Mix is supplied as a 2X pre-mixed formula-tion containing HotStart-IT Taq DNA Polymerase, MgCl2, Ultrapure nucleotides, and SYBR Green I for use in real-time quantitative PCR reactions (qPCR). Simply add DNA template, primers, and water and the reactions are ready for cycling. Separate tubes of the passive reference dyes ROX (for ABI, Stratagene, and other ROX instruments) and fluorescein (for BioRad and other instruments) are included for added convenience.

The master mix has SYBR Green I Dye which detects any double-stranded DNA that accumulates during the amplification process. For carry-over prevention methods, use HotStart-IT SYBR Green PCR Master Mix with UDG (2X), PN 75760 or VeriQuest SYBR Green qPCR Master Mix (2X), PN 75600.

Two passive reference dyesThe SYBR Green I Dye is compatible with any instrument’s standard SYBR filter set (typically, FAM setting) and the separate ROX and Fluorescein Passive Reference Dyes allow normalization of well-to-well variations that may occur independent of the reactions (e.g., pipetting errors, detection system limitations, etc.).

StableRepeated freeze-thaw cycles have no observed effect on performance.

HotStart-IT SYBR Green qPCR Master Mix (2X):The mix combines HotStart-IT Taq DNA Polymerase, SYBR Green I, MgCl2, and Ultrapure nucleotides in a unique buffer formulation. Magnesium and nucleotide concentrations are 5 mM and 0.4 mM each, respectively.

Components:HotStart-IT SYBR Green qPCR Master Mix (2X): 100 reactions 2 x 1.25 ml 500 reactions 12.5 ml25 mM MgCl2ROX Passive Reference DyeFluorescein Passive Reference Dye


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HotStart-IT SYBR Green qPCR Master Mix (2X)



HotStart-IT SYBR Green qPCR Master Mix with Uracil-DNA Glycosylase (UDG or UNG) uses a novel hot start method developed at Affymetrix called primer sequestration. With this method, a protein binds and sequesters primers at lower temperatures making them unavailable for use by Taq DNA Polymerase. Following the initial denaturation step, the protein is inactivated and the primers are released. HotStart-IT SYBR Green qPCR Master Mix with UDG is supplied as a 2X pre-mixed formulation containing HotStart-IT Taq DNA Polymerase, MgCl2, Ultrapure nucleotides with an optimized dUTP to dTTP ratio, heat-labile UDG, and SYBR Green I for use in real-time quantitative PCR reactions (qPCR). Simply add DNA template, primers, and water and the reactions are ready for cycling. Separate tubes of passive reference dyes ROX (for ABI, Stratagene, and other ROX instruments) and fluorescein (for BioRad and other instruments) are included for added convenience.

Since the mix contains dUTP and UDG, carry-over contamination prevention can be performed. A heat-labile version of UDG that can be irreversibly heat-inactivated is used instead of E. coli UDG, which has been shown to exhibit residual activity following PCR reactions(1). The SYBR Green I Dye detects any double-stranded DNA that accumulates during the amplification process. The hot start feature enhances SYBR-based qPCR reactions by reducing primer-dimer formation which increases specificity and sensitivity. HotStart-IT SYBR Green qPCR Master Mix with UDG has excellent sensitivity as it detects fewer than 10 target copies, performs over a linear dynamic range of 7 to 8 orders of mag-nitude, and is compatible with a variety of real-time PCR instruments.

Multiple platform compatibilityThe SYBR Green I Dye is compatible with any instru-ment’s standard SYBR filter (typically, FAM setting) and the separate ROX and Fluorescein Passive Reference Dyes allow normalization of well-to-well variations that may occur independent of the reactions (e.g., pipetting errors, detection system limitations, etc.).


HotStart-IT SYBR Green qPCR Master Mix with UDG Formulation (2X):The mix combines HotStart-IT Taq DNA Polymerase, heat-labile UDG, SYBR Green I, MgCl2, and Ultrapure nucleotides with an optimized dUTP to dTTP ratio in a unique buffer formulation. Magne-sium and nucleotide concentrations are 5 mM and 0.4 mM each, respectively.

Components:HotStart-IT SYBR Green qPCR Master Mix with

UDG (2X) 100 reactions 2 x 1.25 ml 500 reactions 12.5 ml25 mM MgCl2ROX Passive Reference DyeFluorescein Passive Reference Dye


References:1. Thornton, C. G., Hartley, J. L., and Rashtchian, A.

(1992) BioTechniques 13, 180-184.2. Longo, M. C., Berninger, M. S., and Hartley, J. L.

(1990) Gene 93, 125-128.

HotStart-IT SYBR Green qPCR Master Mix with UDG (2X)




VeriQuest Probe qPCR Master Mix is supplied as a 2X pre-mixed formulation containing both ROX and UDG in an optimized buffer for quality results in real-time quantitative PCR assays. The proprietary reaction buffer with optimum MgCl2 concentration is specifically designed for robust probe hybridiza-tion and efficient cleavage of TaqMan probes. Simply add DNA template, primers, probe(s), and water and the reactions are ready for cycling. The master mix contains a chemically-modified Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), and ROX Passive Reference Dye in a proprietary reaction buffer.

Features:�� One-tube master mix with ROX Reference Dye included�� Similar protocols, same probes, and same primers for standard mode amplification�� Consistency over a broad dynamic range: 9 orders of magnitude linear detection range with a correlation coefficient = 0.999�� Exceptional performance with challenging GC-rich regions �� Detect 2 copies of target from genomic DNA and differentiate less than a two-fold difference.�� Contains dUTP and Uracil-DNA Glycosylase (UDG or UNG) for optional carryover contamination prevention.�� Hot start PCR for room temperature reaction setup�� Stable at room temperature for 72 hours in a preassembled reaction�� Validated for use with a variety of 5’ reporter fluorophores and 3’ quenchers, as well as TaqMan MGB containing probes

Reproducible, consistent results while main-taining precision and efficiencyThe convenience of a master mix formulation with-out sacrificing quality. VeriQuest Probe qPCR Master Mix provides consistency over a broad dynamic template concentration range allowing 9 orders of magnitude linear detection (Fig. 1 and 2). High preci-sion in target quantification and discrimination of a 1.33 to 10-fold dilution series with VeriQuest Probe qPCR Master Mix ensure accurate results (Fig. 3).

Enhanced amplification sensitivity and accuracy of low copy number detectionReliable detection from as little as 2 copies of target from genomic DNA and differentiation from less than a two-fold difference (Fig. 3).

Exceptional performance with challenging templatesEven in high GC and AT rich regions VeriQuest Probe qPCR Master Mix offers exceptional performance and high specificity so you have the confidence to verify your gene expression results (Fig. 4).

Ready-to-use, one-tube replacementThe one-tube master mix contains all neces-sary components; chemically-modified Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), and ROX as the pas-sive reference dye, all premixed in a proprietary reaction buffer. A drop-in replacement for use on any leading PCR platform that utilizes ROX.

Passive Reference Dye and UDG includedROX Passive Reference Dye is an inert dye, whose fluorescence does not change during the reaction. This signal normalization is necessary to correct for well-to-well differences that may occur due to artifacts such as pipetting errors or sample evapora-tion. Uracil-DNA Glycosylase (UDG or UNG) and dUTP offer an option for carry-over contamination prevention from previous PCR amplifications. All are at optimized levels so there is no need to adjust concentrations.

Highly stable and easy to work withVeriQuest Probe qPCR Master Mix is stable at room temperature for 72 hours in a pre-assembled reaction and can be stored at 4°C for convenient handling and room temperature reaction set up; no lost time waiting for your master mix to thaw (Figs. 5 and 6). Testing of 10 freeze-thaw cycles showed no loss in master mix performance. Ideal for high-throughput handling.

Consistent performance of single-plex and duplex detectionSingleplex (S) and duplex detection (D) of target genes can be performed using VeriQuest Probe qPCR Master Mix, making this ideal for gene expression verification.

Components:40 reactions: 1 ml200 reactions: 5 ml400 reactions: 2 x 5 ml1,000 reactions: 5 x 5 ml2,000 reactions: 10 x 5 mlPCR Qualified Water

References:1. VanGuilder, H. D., Vrana, K. E., and Freeman, W. M.

(2008) BioTechniques 44 (5), 619-626.2. Longo, M. C., Berninger, M. S., and Hartley, J. L.

(1990) Gene 93, 125-128.3. Tewhey, R. et al. (2009) Nature Biotechnology 27,

1025-1031.

VeriQuest Probe qPCR Master Mix (2X)

Fig. 1. Linear detection range of VeriQuest Probe qPCR Master Mix. Real-time amplification plot from a 10-fold dilution series of a GAPDH synthetic target with starting amounts of 1010 copies amplified in four replicate reactions using the ABI 7500 PCR System and GAPDH primer-probe set (FAM-BHQ®-1).

Fig. 2. Inset for linear detection range. Amplification plot from a 10-fold dilution series.

Fig. 3. High precision in target quantification with VeriQuest Probe qPCR Master Mix. Amplification plot from real-time PCR for a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNAs reverse-transcribed from HeLa total RNA.



VeriQuest Probe qPCR Master Mix (2X) continued...

Fig. 4. Amplification of GC-rich target with VeriQuest Probe qPCR Master Mix. qPCR standard curves for a 10-fold dilution series of 10 ng to 10 pg human male genomic DNA amplified in four replicate reactions with VeriQuest Probe qPCR Master Mix, ABI TaqMan® Gene Expression Master Mix and ABI TaqMan Universal Master Mix II using the ABI 7500 PCR System for detection of a 71.2% GC-target amplicon (PROC, NT_022135.16)(3).

Fig. 6. Inset defining Ct levels of 1 ng and 10 pg of cDNAs reverse transcribed from HeLa total RNA. GAPDH was detect-ed from preassembled PCR reactions incubated at room temperature for 48 and 72 hours.

Fig. 5. VeriQuest Probe qPCR Master Mix stability for reli-able performance of high-throughput handling. Pre-assembled PCR reaction stability after 48 and 72 hour incubations at room temperature from quantification of GAPDH cDNAs.

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VeriQuest Fast Probe qPCR Master Mix (2X)75680 100 reactions


VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X)75685 100 reactions






100 ml






VeriQuest Probe qPCR Master Mix, No Reference Dye is supplied as a 2X pre-mixed formulation con-taining UDG in an optimized buffer for quality results in real-time quantitative PCR assays. The proprietary reaction buffer with optimum MgCl2 concentration is specifically designed for robust probe hybridization and efficient cleavage of TaqMan probes. Simply add DNA template, primers, probe(s), and water, and the reactions are ready for cycling.

Features:�� Enhanced amplification sensitivity and accuracy of low copy number detection�� Reproducible, consistent results while maintaining precision and efficiency�� Exceptional performance with challenging templates�� Ready-to-use, one-tube replacement�� UDG included�� Highly stable and easy to work with�� Ample detection of up to 4 different target genes

Enhanced sensitivity on amplification for low copy number detectionReliable detection from as little as two copies of target from genomic DNA and differentiation from less than a two-fold difference.

Reproducible, consistent results while main-taining precision and efficiencyThe convenience of a master mix formulation with-out sacrificing quality. VeriQuest Probe qPCR Master Mix, No Reference Dye provides consistency over a broad dynamic template concentration range, al-lowing 9 orders of magnitude linear detection. High precision in target quantification and discrimination of a 1.33 to 10-fold dilution series with VeriQuest Probe qPCR Master Mix, No Reference Dye ensures accurate results.

Exceptional performance with challenging templatesEven in high GC and AT rich regions, VeriQuest Probe qPCR Master Mix, No Reference Dye offers excep-tional performance and high specificity so you have the confidence to verify your gene expression results.

Ready-to-use, one-tube replacementThe one-tube master mix contains all necessary com-ponents: chemically-modified Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, and Uracil-DNA Glycosylase (UDG or UNG) in a proprietary reaction buffer. Simply add your DNA template, probe, primers, and water, and you can begin your qPCR reaction. For use in your new or existing protocols, on any leading PCR plat-form that does not require a passive reference dye, providing easy transition from other master mixes. Examples of PCR platforms include: Eppendorf Mastercycler®, Helixis Pixo™, Qiagen Rotor-Gene™ Q, Roche LightCycler® 480/1536, TaKaRa TP-800, Bio-Rad CFX96/CFX384, Bio-Rad Opticon 2™, Bio-Rad Chromo 4™, Cepheid Smart Cycler®, and Corbett Rotor-Gene™.

UDG included for carry-over contamination preventionUracil-DNA Glycosylase (UDG or UNG) and dUTP offer an option for carry-over contamination preven-tion from previous PCR amplifications. Both are at optimized levels so there is no need to adjust concentrations.

Highly stable and easy to work withVeriQuest Probe qPCR Master Mix, No Reference Dye is stable at room temperature for 72 hours in a preassembled reaction. This mix can be stored at 4°C for convenient handling and room temperature reaction set up, with no time lost waiting for your master mix to thaw. Testing of 10 freeze thaw cycles showed no loss in master mix performance, making it ideal for high-throughput handling.

Ample detection of up to 2 different target genesSingle-plex (S) and duplex detection (D) of target genes is possible with VeriQuest Probe qPCR Master Mix, No Reference Dye for gene expression verification.

Components:40 reactions: 1 ml200 reactions: 5 ml400 reactions: 2 x 5 ml1,000 reactions: 5 x 5 ml2,000 reactions: 10 x 5 mlPCR Qualified Water

Storage:Shipped on dry ice. Store at -20°C for long-term storage. Store at 4-8°C for short-term storage (≤3 months).

VeriQuest Probe qPCR Master Mix, No Reference Dye (2X)

Real-time amplification plot and standard curve from a 10-fold dilution series of a synthetic target with starting amounts of 1010 copies ampli-fied in four replicate reactions using the Bio-Rad MyiQ Real-Time PCR System and GAPDH primer-probe set (Fam-BHQ®-1).

Linear detection range of VeriQuest Probe qPCR Master Mix, No Reference Dye



VeriQuest Fast Probe qPCR Master Mix (2X)

75680 (20 µl reaction volume)100 reactions (1 ml)500 reactions (5 ml)

1,000 reactions (2 x 5 ml)2,500 reactions (5 x 5 ml)

5,000 reactions (10 x 5 ml)

Features:�� Uses fast mode thermal cycling conditions for results in one-third of the time when compared to standard run protocols.�� One-tube master mix with ROX Reference Dye included – just add template, primers and water �� Exceptional performance with challenging GC-rich regions�� Contains dUTP and Uracil-DNA Glycosylase (UDG or UNG) for carryover contamination prevention.�� Hot start PCR for room temperature reaction setup�� Sensitivity and precision with limited targets

VeriQuest Fast Probe qPCR Master Mix (2X) is a ready-to-use master mix optimized for TaqMan® probe detection on all instruments using fast mode cycling protocols that utilize ROX passive reference dye. The 2X ready-to-use master mix contains hot start VeriQuest Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), and ROX Passive Reference Dye in a proprietary reaction buffer. The hot start Taq polymerase has no polymerase activity prior to the initial heat activation step, which allows reaction assembly at room temperature as well as higher specificity and sensitivity. Since the mix contains dUTP and UDG, carryover contamination prevention can be performed prior to amplification. VeriQuest Fast Probe qPCR Master Mix offers the

same sensitivity and specificity found with our stan-dard mode mixes with results in a fraction of the time.

VeriQuest Fast Probe qPCR Master Mix shows excep-tional efficiency and high specificity on challenging templates such as high GC- and AT-rich regions. The optimized formulation ensures no sacrifice in quality for increased speed. Performance comparisons highlight the sensitivity and reliable performance of this platform-independent solution.The sensitivity of the master mix allows for discrimination from a 1.33-fold difference in gene target amount detected. In Fig. 1, a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNA reverse-transcribed from HeLa total RNA was amplified with an efficiency of >98.8%.

VeriQuest Fast Probe qPCR Master Mix is highly stable and easy to work with. It remains stable at

room temperature for 72 hours in a pre-assembled reaction and can be stored at 4°C for convenient handling. The mix also allows for room tempera-ture reaction set-up. The speed and stability make VeriQuest Fast Probe qPCR Master Mix ideal for high-throughput handling.



Linear detection range of VeriQuest Fast Probe qPCR Master Mix. Amplification plot and standard curve from real-time PCR for a dilution series of a synthetic target with starting amounts of 1010 cop-ies amplified in four replicate reactions using the ABI 7500 Real-Time PCR System and GAPDH primers in fast mode with 5 minutes activa-tion at 95°C. The amplification process was linear over eight orders of magnitude.


Fig. 2. High sensitivity and precision in limited target quantification. Amplification plot (bottom) and standard curve (top) from real-time PCR for a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNA reverse-transcribed from HeLa total RNA.

Cyclophilin

10 4 3 2 1 ng RNA

Kras

10 4 3 2 1 ng RNA

Cyclophiliny = -3.3841x + 24.142

EPCR = 97.5%R² = 0.9977

Krasy = -3.3502x + 31.829

EPCR = 98.8%R² = 0.9973

20

22

24

26

28

30

32

34

-0.2 0 0.2 0.4 0.6 0.8 1 1.2

Ct

log input

Cyclophilin Kras

Fig. 3. VeriQuest Fast Probe qPCR Master Mix stability. GAPDH was detected from pre-assembled PCR reactions incubated at room temperature for 72 hours. Results were compared to the freshly prepared mix (0 hours).

14

19

24

29

34

39

-3.0 -2.0 -1.0 0.0 1.0 2.0 3.0

Ct

0 hours 72 hours

0 hours y = -3.4119x + 24.224

EPCR = 96.7%R² = 0.9995

72 hours y = -3.3581x + 24.134

EPCR= 98.5%R² = 0.9996



500 reactions (5 ml) 1,000 reactions (2 x 5 ml) 2,500 reactions (5 x 5 ml) 5,000 reactions (10 x 5 ml)

Features:�� Uses fast mode thermal cycling conditions for results in one-third of the time when compared to standard run protocols.�� One-tube master mix – just add template, primers, and water �� Exceptional performance with challenging GC-rich regions�� Contains dUTP and Uracil-DNA Glycosylase (UDG or UNG) for carryover contamination prevention.�� Hot start PCR for room temperature reaction setup�� Sensitivity and precision with limited targets

VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X) is a ready-to-use master mix optimized for TaqMan® probe detection on all instruments using fast mode cycling protocols that do not require use of a passive reference dye. The 2X ready-to-use master mix contains hot start VeriQuest Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, and Uracil-DNA Glycosylase (UDG or UNG) in a proprietary reaction buffer. The hot start Taq polymerase has no polymerase activity prior to the initial heat activation step, which allows reaction assembly at room tem-perature as well as higher specificity and sensitivity. Since the mix contains dUTP and UDG, carryover contamination prevention can be performed prior to amplification. VeriQuest Fast Probe qPCR Master

Mix, No Reference Dye offers the same sensitivity and specificity found with our standard mode mixes with results in a fraction of the time.

VeriQuest Fast Probe qPCR Master Mix, No Refer-ence Dye shows exceptional efficiency and high specificity on challenging templates such as high GC- and AT-rich regions The optimized formulation ensures no sacrifice in quality for increased speed. Performance comparisons highlight the sensitivity and reliable performance of this platform-indepen-dent solution.The sensitivity of the master mix allows for discrimination from a 1.33-fold difference in gene target amount detected. In Fig. 1, a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNA reverse-transcribed from HeLa total RNA was ampli-fied with an efficiency of >98.8%. VeriQuest Fast Probe qPCR Master Mix, No Reference Dye is highly

stable and easy to work with. It remains stable at room temperature for 72 hours in a pre-assembled reaction and can be stored at 4°C for convenient handling. The mix also allows for room temperature reaction set-up. The speed and stability make Veri-Quest Fast Probe qPCR Master Mix, No Reference Dye ideal for high-throughput handling.



VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X)


Linear detection range of VeriQuest Fast Probe qPCR Master Mix. Amplification plot and standard curve from real-time PCR for a dilution series of a synthetic target with starting amounts of 1010 cop-ies amplified in four replicate reactions using the ABI 7500 Real-Time PCR System and GAPDH primers in fast mode with 5 minutes activa-tion at 95°C. The amplification process was linear over eight orders of magnitude.

Fig. 2. High sensitivity and precision in limited target quantification. Amplification plot (bottom) and standard curve (top) from real-time PCR for a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNA reverse-transcribed from HeLa total RNA.

Cyclophilin

10 4 3 2 1 ng RNA

Kras

10 4 3 2 1 ng RNA

Cyclophiliny = -3.3841x + 24.142

EPCR = 97.5%R² = 0.9977

Krasy = -3.3502x + 31.829

EPCR = 98.8%R² = 0.9973

20

22

24

26

28

30

32

34

-0.2 0 0.2 0.4 0.6 0.8 1 1.2

Ct

log input

Cyclophilin Kras

Fig. 3. VeriQuest Fast Probe qPCR Master Mix stability. GAPDH was detected from pre-assembled PCR reactions incubated at room temperature for 72 hours. Results were compared to the freshly prepared mix (0 hours).

14

19

24

29

34

39

-3.0 -2.0 -1.0 0.0 1.0 2.0 3.0

Ct

0 hours 72 hours

0 hours y = -3.4119x + 24.224

EPCR = 96.7%R² = 0.9995

72 hours y = -3.3581x + 24.134

EPCR= 98.5%R² = 0.9996



HotStart-IT Probe qPCR Master Mix uses a novel hot start method developed at Affymetrix called primer sequestration. With this method, a protein binds and sequesters primers at lower temperatures mak-ing them unavailable for use by Taq DNA Polym-erase. Following the initial denaturation step, the protein is inactivated and the primers are released. HotStart-IT Probe qPCR Master Mix is supplied as a 2X pre-mixed formulation containing HotStart-IT Taq DNA Polymerase, MgCl2, and Ultrapure nucleo-tides for use in real-time quantitative PCR reactions (qPCR) with fluorescent probes. Simply add DNA template, primers, probe(s), and water and the reac-tions are ready for cycling. A separate tube of ROX passive reference dye (for ABI, Stratagene, and other instruments) is included for added convenience.

The master mix is formulated for use with fluo-rescent probes such as TaqMan Probes, Molecular Beacons, and others(1-2). Since fluorescent probes are designed to hybridize to the target of interest, detection specificity is greatly increased relative to non-specific dsDNA binding dyes such as SYBR Green I. The Taq DNA Polymerase used in the master mix has the 5'→3' exonuclease activity necessary for efficient removal of the 5'-fluorophore from the 3'-quencher in TaqMan probes. HotStart-IT Probe qPCR Master Mix has excellent sensitivity as it detects fewer than 10 target copies, performs over a broad, linear dynamic range of 7 to 8 orders of magnitude, and is compatible with a variety of real-time PCR instruments. The mix does not have dUTP in place of dTTP and is incompatible with carry-over contamination prevention methods using Uracil-DNA Glycosylase (UDG or UNG). For carry-over prevention methods, use HotStart-IT Probe qPCR Master Mix with UDG (2X), PN 75764 or VeriQuest Probe qPCR Master Mix (2X), PN 75650.


HotStart-IT Probe qPCR Master Mix (2X):The mix combines HotStart-IT Taq DNA Polymerase (with 5'→3' exonuclease activity), MgCl2, and Ultrapure nucleotides in a unique buffer formulation. Magnesium and nucleotide concentrations are 6 mM and 0.4 mM each, respectively.

Components:HotStart-IT Probe qPCR Master Mix (2X) 100 reactions 2 x 1.25 ml 500 reactions 12.5 ml25 mM MgCl2ROX Passive Reference Dye


References:1. Livak, K. J., Flood, S. J., Marmaro, J., Giusti, W., and

Deetz, K. (1995) PCR Methods Appl. 4, 357-362.2. Tyagi, S., and Kramer, F. R. (1996) Nat. Biotechnol.

14, 303-308.

HotStart-IT Probe qPCR Master Mix (2X)


HotStart-IT Probe qPCR Master Mix with Uracil-DNA Glycosylase (UDG or UNG) uses a novel hot start method developed at Affymetrix called primer sequestration. With this method, a protein binds and sequesters primers at lower temperatures making them unavailable for use by Taq DNA Polymerase. Following the initial denaturation step, the protein is inactivated and the primers are released. HotStart-IT Probe qPCR Master Mix with UDG is supplied as a 2X pre-mixed formulation containing HotStart-IT Taq DNA Polymerase, MgCl2, Ultrapure nucleotides with an optimized dUTP to dTTP ratio, and heat-labile UDG for use in real-time quantitative PCR reactions (qPCR) with fluorescent probes. Simply add DNA template, primers, probe(s), and water and the reac-tions are ready for cycling. A separate tube of ROX passive reference dye (for ABI, Stratagene, and other instruments) is included for added convenience.

Since the mix contains dUTP and UDG, carry-over contamination prevention can be performed. A heat-labile version of UDG that can be irreversibly heat-inactivated is used instead of E. coli UDG, which has been shown to exhibit residual activity

following PCR reactions(1). The mix is formulated for use with fluorescent probes such as TaqMan Probes, Molecular Beacons, and others(3-4). Since fluorescent probes are designed to hybridize to the target of interest, detection specificity is greatly increased relative to non-specific dsDNA binding dyes such as SYBR Green I. The Taq DNA Polymerase used in the master mix has the 5'→3' exonuclease activity necessary for efficient removal of the 5'-fluorophore from the 3'-quencher in TaqMan probes. HotStart-IT Probe qPCR Master Mix with UDG has excellent sensitivity as it detects fewer than 10 target copies, performs over a broad, linear dynamic range of 7 to 8 orders of magnitude, and is compatible with a vari-ety of real-time PCR instruments.

Multiple platform compatibilitySpecialized buffer with optimal MgCl2 concentration and dUTP to dTTP ratio performs well on a variety of platforms. Also, the separate tube of ROX Passive Reference Dye allows normalization of well-to-well variations that may occur independent of the reactions (e.g., pipetting errors, detection system limitations, etc.).


HotStart-IT Probe qPCR Master Mix with UDG (2X):The mix combines HotStart-IT Taq DNA Polymerase (with 5'→3' exonuclease activity), heat-labile UDG, MgCl2, and Ultrapure nucleotides with an optimized dUTP to dTTP ratio in a unique buffer formulation. Magnesium and nucleotide concentrations are 6 mM and 0.4 mM each, respectively.

Components:HotStart-IT Probe qPCR Master Mix with UDG (2X) 100 reactions 2 x 1.25 ml 500 reactions 12.5 ml25 mM MgCl2ROX Passive Reference Dye


References:1. Thornton, C. G., Hartley, J. L., and Rashtchian, A.

(1992) BioTechniques 13, 180-184.2. Longo, M. C., Berninger, M. S., and Hartley, J. L.

(1990) Gene 93, 125-128.3. Livak, K. J., Flood, S. J., Marmaro, J., Giusti, W., and

Deetz, K. (1995) PCR Methods Appl. 4, 357-362.4. Tyagi, S., and Kramer, F. R. (1996) Nat. Biotechnol.

14, 303-308.

HotStart-IT Probe qPCR Master Mix with UDG (2X)



75780 50 reactions (20 µl)

The First-Strand cDNA Synthesis Kit for Real-Time PCR is optimized for reverse transcription of RNA and produces first-strand cDNA template suitable for real-time PCR.

�� Robust performance: Optimized to reliably generate cDNA from full-length transcripts longer than 12 kb.�� Sensitive: Sensitive and reliable real-time RT-PCR analysis of multiple user-defined targets from as little as 10 pg of starting total RNA.�� Consistent performance: Optimized for generating first-strand cDNA to be used in real-time PCR. HeLa total RNA and primers for qPCR are included as a positive control.�� Versatile: Three different priming strategies are provided to meet experimental needs. Mixed priming strategy overcomes 5' and/or 3' end bias associated with typical oligo(dT)n or random hexamer priming strategies.

Applications:Conversion of RNA into single-stranded cDNA involves a complex, enzyme-catalyzed reaction known as reverse transcription (RT). RT followed by real-time or quantitative PCR (real-time RT-PCR or qPCR) amplification is the most sensitive technique for mRNA detection and quantification currently available(1).

This kit is specifically designed to convert RNA into first-strand cDNA that is suitable for real-time PCR applications.

First-strand cDNA generated by this kit is also suitable for end-point PCR. RT-PCR can be used in a variety of applications, such as qualitative and quantitative analyses of cellular RNAs, characteriza-tion of RNA splice variants, and the generation and cloning of cDNAs.

Mixed priming strategy eliminates end-biasThis kit includes an optimized Primer Mix which results in the generation of first-strand cDNAs from an entire transcript without the end-bias observed with typical oligo(dT)n or random hexamer primers (Fig. 1). This mixed primer strategy overcomes variability in real-time PCR gene expression analysis that can result from using different individual primers(2). For convenience, anchored oligo(dT)23VN and random hexamer primers are also included.

Robust and sensitive performanceThis kit has been designed to routinely generate cDNAs from transcripts longer than 12 kb (Fig. 2). Consistent results are obtained using any of the three priming strategies provided with this kit. Reverse transcription can be performed with as little as 10 pg of total RNA, allowing sensitive down-stream analysis (Fig. 3).

Kit components:10X RT Buffer10 mM dNTPsRNase Inhibitor (10 units/µl)RNase-Free, DEPC-Treated WaterM-MLV RTAnchored Oligo(dT)23VN (50 µM)Random Hexamers (75 µM )10X Primer MixHeLa Total RNA (100 ng/µl)Control Primer Mix for qPCR

References:1. Bustin, S., Benes, V., Nolan, T., and Pfaffl, M.

(2005) J. Mol. Endocrinology 34, 597-601.2. Ståhlberg, A., Håkansson, J., Xian, X., Semb, H.,

and Kubista, M. (2004) Clinical Chem. 50(3), 509-515.

First-Strand cDNA Synthesis Kit for Real-Time PCR

Fig. 1. The Primer Mix solution greatly reduces bias for sequences near the 5' and/or 3' ends of cDNAs produced. The β-glucuronidase (GUSB) mRNA was reverse transcribed from 100 ng of HeLa cell total RNA using (A) Random Hexamers, (B) Anchored Oligo(dT)23VN, or (C) the Primer Mix. Amplicons located near the 5' end (red) or 3' end (green) of the GUSB transcript were amplified by real-time PCR (on an ABI 7500 Fast instrument) using 1 µl of each reverse transcription reaction in 20 µl real-time PCR reactions (in duplicate) and HotStart-IT SYBR Green qPCR Master Mix (PN 75762).

Fig. 2. Assay flexibility and consistent priming strategies. The First-Strand cDNA Synthesis Kit for Real-Time PCR has been opti-mized for assay flexibility in carrying out reverse transcription using various priming strategies and diverse RNA templates. (A) Clathrin (NM_004859) and (B) Utrophin (NM_007124) amplicons were amplified using gene specific primers designed for real-time PCR (brown arrows). Reverse transcription was performed using HeLa cell total RNA (100 ng) and three different priming strategies: Anchored Oligo dT23VN primer (red), Random Hexamers (blue), and Primer Mix (green). 1 µl of each reverse transcription reaction was then used in 20 µl real-time PCR reactions on an ABI 7500 Fast instrument using HotStart-IT SYBR Green qPCR Master Mix (PN 75762).

Fig. 3. Assay sensitivity and dynamic range. Various amounts of HeLa cell total RNA, ranging from 850 ng down to 10 pg, were reverse transcribed using anchored oligo(dT)23VN as the primer. 1 µl of each reverse transcription reaction was then used in 20 µl real-time PCR reactions (in duplicate) to amplify a 122 bp GAPDH amplicon (on an ABI 7500 Fast instrument), using HotStart-IT SYBR Green qPCR Master Mix (PN 75762). GAPDH amplification and linear correlation curve above show the wide dynamic range and sensitivity of the First-Strand cDNA Synthesis Kit for Real-Time PCR.

PCR Tools | Real-Time Reverse Transcription PCR (qRT-PCR)


VeriQuest SYBR Green One-Step qRT-PCR Master Mix (2X)


200 reactions (5 ml)

Features:�� Reproducible, consistent results while maintaining precision and efficiency�� Multiple platform compatibility with all real-time master mixes�� One-step, sequential reaction format for minimum hands on time�� Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contamination�� Exceptional stability even after 10 freeze-thaw cycles

VeriQuest SYBR Green One-Step qRT-PCR Master Mix is a one-step, ready-to-use master mix for RNA quantification with SYBR Green detection. The optimized buffer formulation offers specificity, ro-bustness, and reliability. The RT-PCR process converts and amplifies single-stranded RNA template yielding double-stranded DNA product. One-Step RT-PCR uses gene specific primers, designed to match RNA/cDNA targets, in a single-tube/plate, one-step reac-tion. This approach offers tremendous convenience when applied to analysis of single targets from mul-tiple RNA samples. Also, it minimizes the possibility of introducing contaminants into reactions between the RT and PCR steps, since both steps are carried out sequentially without opening the reaction tubes/plates between the steps(1-5).

Reproducible, consistent results while main-taining precision and efficiencyThis master mix provides the convenience of a master mix formulation without sacrificing quality. VeriQuest SYBR Green One-Step qRT-PCR Master Mix exhibits excellent sensitivity, as it can detect fewer than 10 target copies and performs over a broad, linear dynamic range of 8 orders of magni-tude (Figs. 1 and 2).

Multiple platform compatibility with all real-time master mixesThis master mix contains ROX passive reference dye in an optimized concentration for use on all instru-ments that require ROX dye for signal normalization. ROX Passive Reference Dye is an inert dye, whose fluorescence does not change during the reaction. This signal normalization is necessary to correct for well-to-well differences that may occur due to arti-facts such as pipetting errors or sample evaporation. VeriQuest SYBR Green OneStep qRT-PCR Master Mix is also available for instruments that require fluorescein. To determine which master mix is appro-priate for your thermal cycler, refer to our instrument compatibility guide.

Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contaminationThe 2X master mix contains 100X RT Enzyme Mix and 2X qRT-PCR Mix for RNA quantification with SYBR Green. The 100X RT Enzyme Mix is a blend of reverse transcriptase and RNase Inhibitor. The 2X qRT-PCR Mix contains hot start VeriQuest Taq DNA Polymerase, ultrapure nucleotides, SYBR Green I and ROX Passive Reference Dye in an optimized buffer formulation for quantitative, real-time PCR detection with SYBR Green.

Exceptional stability even after 10 freeze-thaw cyclesVeriQuest SYBR Green One-Step qRT-PCR Master Mix is stable and easy to work with. Performance testing of 10 freeze thaw cycles showed no loss in master mix performance, making this ideal for high-throughput handling.

Components:100X RT Enzyme Mix for SYBR Green Assay2X qRT-PCR Mix with SYBR Green 40 reactions 1 ml200 reactions 5 ml


References:1. Goblet, C., Prost, E., and Whalen, R. G. (1989)

Nucleic Acids Res. 17, 2144.2. Sambrook, J. and Russell, D. W. (2001) “Molecular

Cloning: A Laboratory Manual,” Cold Spring Harbor Laboratory Press, 8.46-8.53.

3. Sellner, L. N., Coelen, R. J., and MacKenzie, J. S. (1992) Nucleic Acids Res. 20, 1487-1490.

4. Roth, M. J., Tanese, N., and Goff, S. P. (1985) J. Biol. Chem. 260, 9326-9335.

5. Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., and Erlich, H. A. (1988) Science 239, 487-491.

Fig. 2. Melt Curve analysis for VeriQuest SYBR Green One-Step qRT-PCR Master Mix.

Fig. 1. Consistent results over an extremely broad dynamic range. Linear detection range of VeriQuest SYBR Green One-Step qRT-PCR Master Mix. RNA from human muscle cells was reverse tran-scribed and amplified using 18S primers. Triplicate reactions were run on an ABI 7500 Real-Time PCR System



VeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein (2X)



Features:�� Reproducible, consistent results while maintaining precision and efficiency�� Multiple platform compatibility with all real-time master mixes�� One-step, sequential reaction format for minimum hands on time�� Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contamination�� Exceptional stability even after 10 freeze-thaw cycles

VeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein is a one-step, ready-to-use master mix for RNA quantification with SYBR Green detection. The optimized buffer formulation offers specificity, robustness, and reliability. The RT-PCR process converts and amplifies single-stranded RNA template yielding double-stranded DNA product. One-Step RT-PCR uses gene specific primers, de-signed to match RNA/cDNA targets, in a single-tube/plate, one-step reaction.

This approach offers tremendous convenience when applied to analysis of single targets from multiple RNA samples. Also, it minimizes the possibility of introducing contaminants into reactions between the RT and PCR steps, since both steps are carried out sequentially without opening the reaction tubes/plates between the steps(1-5).

Reproducible, consistent results while main-taining precision and efficiencyThis master mix provides the convenience of a master mix formulation without sacrificing quality. VeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein exhibits excellent sensitivity, as it can detect fewer than 10 target copies and performs over a broad, linear dynamic range of 8 orders of magnitude (Figs. 1 and 2).

Multiple platform compatibility with all real-time master mixesThis master mix contains fluorescein passive refer-ence dye in an optimized concentration for use on instruments that require fluorescein dye for signal normalization (i.e., BioRad iCycler, MyiQ, iQ5). Fluo-rescein Passive Reference Dye is an inert dye, whose fluorescence does not change during the reaction. This signal normalization is necessary to correct for well-to-well differences that may occur due to arti-facts such as pipetting errors or sample evaporation. VeriQuest SYBR Green One-Step qRT-PCR Master Mix is also available for instruments that require ROX reference dye. To determine which master mix is appropriate for your thermal cycler, refer to our instrument compatibility guide.

Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contaminationThe 2X master mix contains 100X RT Enzyme Mix and 2X qRT-PCR Mix for RNA quantification with SYBR Green. The 100X RT Enzyme Mix is a blend of reverse transcriptase and RNase Inhibitor. The 2X qRT-PCR Mix contains chemically-modified VeriQuest Taq DNA Polymerase, ultrapure nucleotides, SYBR Green I, and Fluorescein Passive Reference Dye in optimized buffer formulation for quantitative, real-time PCR detection with SYBR Green.

Exceptional stability even after 10 freeze-thaw cyclesVeriQuest SYBR Green One-Step qRT-PCR Master Mix with Fluorescein is stable and easy to work with. Performance testing of 10 freeze thaw cycles showed no loss in master mix performance, making this ideal for high-throughput handling.

Components:100X RT Enzyme Mix for SYBR Green Assay2X qRT-PCR Mix with SYBR Green 40 reactions 1 ml200 reactions 5 ml









Fig. 2. Melt Curve analysis for VeriQuest SYBR Green One-Step qRT-PCR Master Mix.

Fig. 1. Consistent results over an extremely broad dynamic range. Linear detection range of VeriQuest SYBR Green One-Step qRT-PCR Master Mix. RNA from human muscle cells was reverse tran-scribed and amplified using 18S primers. Triplicate reactions were run on an ABI 7500 Real-Time PCR System


VeriQuest Probe One-Step qRT-PCR Master Mix (2X)



Features:�� Reproducible, consistent results while maintaining precision and efficiency�� Multiple platform compatibility with all real-time master mixes�� One-step, sequential reaction format for minimum hands on time�� Easily perform analysis of single or duplex targets from multiple RNA samples�� Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contamination�� Exceptional stability even after 10 freeze-thaw cycles

VeriQuest Probe One-Step qRT-PCR Master Mix is a one-tube, one-step, ready-to-use master mix for RNA quantification with TaqMan® probe detection. The optimized buffer formulation offers specificity, ro-bustness, and reliability. The RT-PCR process converts and amplifies single-stranded RNA template yielding double-stranded DNA product. One-Step RT-PCR uses gene specific primers, designed to match RNA/cDNA targets, in a single-tube/plate, one-step reac-tion. This approach offers tremendous convenience when applied to analysis of single targets from mul-tiple RNA samples. Also, it minimizes the possibility of introducing contaminants into reactions between the RT and PCR steps, since both steps are carried out sequentially without opening the reaction tubes/plates between the steps(1-5).

Reproducible, consistent results while main-taining precision and efficiencyThis master mix provides the convenience of a master mix formulation without sacrificing quality. VeriQuest Probe One-Step qRT-PCR Master Mix ex-hibits excellent sensitivity, as it can detect fewer than 10 target copies and performs over a broad, linear dynamic range of 8 orders of magnitude (Fig. 1).

Multiple platform compatibility with all real-time master mixesThis master mix contains ROX passive reference dye in an optimized concentration for use on all instru-ments that require ROX dye for signal normalization. ROX Passive Reference Dye is an inert dye, whose fluorescence does not change during the reaction. This signal normalization is necessary to correct for well-to-well differences that may occur due to arti-facts such as pipetting errors or sample evaporation.

Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contaminationThe 2X master mix contains reverse transcriptase, RNase inhibitor, our chemically modified VeriQuest Taq DNA polymerase, ROX passive reference dye, and ultrapure nucleotides all in an optimized buffer composition. Simply add your RNA template, probe, primers, and water, and you can begin your qRT-PCR reaction. For use in your new or existing protocols, on any leading real-time qPCR platform that utilizes ROX.

Exceptional stability even after 10 freeze-thaw cyclesVeriQuest Probe One-Step qRT-PCR Master Mix is stable and easy to work with. Performance testing of 10 freeze thaw cycles showed no loss in master mix performance, making this ideal for high-throughput handling.

Components:40 reactions 1 ml200 reactions 5 ml



Nucleic Acids Res. 17, 2144. 2. Sambrook, J. and Russell, D. W. (2001) “Molecular






Fig. 1. Consistent results over an extremely broad dynamic range. Linear detection range of VeriQuest Probe One-Step qRT-PCR Master Mix. RNA from human muscle cells was reverse transcribed and amplified using 18S primers. Triplicate reactions were run on an ABI 7500 Real-Time PCR System


VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye (2X)



Features:�� Reproducible, consistent results while maintaining precision and efficiency�� Multiple platform compatibility with all real-time master mixes�� One-step, sequential reaction format for minimum hands on time�� Easily perform analysis of single or duplex targets from multiple RNA samples�� Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contamination�� Exceptional stability even after 10 freeze-thaw cycles

VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye is a one-tube, one-step, ready-to-use master mix for RNA quantification with TaqMan® probe detection. The optimized buffer formulation of-fers specificity, robustness, and reliability. The RT-PCR process converts and amplifies single-stranded RNA template yielding double-stranded DNA product.

One-Step RT-PCR uses gene specific primers, designed to match RNA/cDNA targets, in a single-tube/plate, one-step reaction. This approach offers tremendous convenience when applied to analysis of single targets from multiple RNA samples. Also, it minimizes the possibility of introducing contami-nants into reactions between the RT and PCR steps, since both steps are carried out sequentially without opening the reaction tubes/plates between the steps(1-5).

Reproducible, consistent results while main-taining precision and efficiencyThis master mix provides the convenience of a master mix formulation without sacrificing quality. VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye exhibits excellent sensitivity, as it can detect fewer than 10 target copies and performs over a broad, linear dynamic range of 8 orders of magnitude (Fig. 1). Product comparisons show opti-mal results in head-to-head evaluations (Fig. 2).

Multiple platform compatibility with all real-time master mixesThis master mix does not contain a passive reference dye, making it useful for thermo cyclers that do not

require the use of a dye for signal normalization (i.e., LightCycler®, RotorGene, Opticon®). To determine which master mix is appropriate for your thermal cycler, refer to our instrument compatibility guide.

Easily perform analysis of single, duplex, or triplex targets from multiple RNA samplesDuplex detection of target RNA using VeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye can be performed with the same sensitivity and specificity as single-plex reactions.

Pre-mixed format with VeriQuest qRT-PCR mixes reduces potential contaminationThe 2X master mix contains reverse transcriptase, RNase inhibitor, our chemically modified VeriQuest Taq DNA Polymerase, and ultrapure nucleotides all in an optimized buffer composition. Simply add your RNA template, probe, primers, and water, and you can begin your qRT-PCR reaction. For use in your new or existing protocols, or on any leading real-time qPCR platform that does not require the use of a reference dye.

Exceptional stability even after 10 freeze-thaw cyclesVeriQuest Probe One-Step qRT-PCR Master Mix, No Reference Dye is stable and easy to work with. Per-formance testing of 10 freeze thaw cycles showed no loss in master mix performance making this ideal for high-throughput handling.

Components:40 reactions 1 ml200 reactions 5 ml









Fig. 1. Consistent results over an extremely broad dynamic range. Linear detection range of VeriQuest Probe One-Step qRT-PCR Master Mix. RNA from human muscle cells was reverse transcribed and amplified using 18S primers. Triplicate reactions were run on an ABI 7500 Real-Time PCR System

13.0

19.8 23.4

26.8

30.4 33.2

14.0

20.9

24.4 27.8

30.7 32.9

13.3

20.0

23.6 26.9

30.5 33.5

0

5

10

15

20

25

30

35

40

100 ng 10 ng 1 ng 100 pg 10 pg 1 pg 100 fg

Ct

HeLa Total RNA

ABI TaqMan Fast Virus 1-Step

ABI TaqMan RNA-to-CT™ 1-Step

VeriQuest One-Step Probe

16.5 17.5 16.6

slope EPCR R2

ABI TaqMan Fast Virus 1-Step -3.4087 96.50% 0.9994

ABI TaqMan RNA-to-CT 1-Step -3.2142 104.70% 0.9953

VeriQuest One-Step Probe -3.4057 96.62% 0.9998

Fig. 2. Exceptional results with performance comparisons. Product comparison—VeriQuest Probe One-Step qRT-PCR Master Mix. Ct values, R2, and PCR efficiency from real-time PCR for a 10-fold dilution series of 100 ng to 100 fg HeLa total RNA amplified in duplicate reac-tions with VeriQuest Probe One-Step qRT-PCR Master Mix, ABI TaqMan Fast Virus 1-Step Master Mix, and ABI TaqMan RNA-to-CT 1-Step Kit using the ABI 7500 Fast Real-Time PCR System and GAPDH primers and probe (Fam-BHQ) in standard mode with 15 minute RT reaction at 50°C.



Description:HotStart-IT SYBR Green One-Step qRT-PCR Master Mix Kit provides optimal performance and maximum convenience for real-time, quantitative analysis of RNA templates in a single reaction format. The RT-PCR process converts and amplifies single-stranded RNA template yielding double-stranded DNA product. One-step RT-PCR uses gene specific primers, designed to match RNA/cDNA targets, in a single-tube/plate, one-step reaction. This approach offers tremendous convenience when applied to analysis of single targets from multiple RNA samples. Also, it minimizes the possibility of introducing con-taminants into reactions between the RT and PCR steps, since both steps are carried out sequentially without opening the reaction tubes/plates between the steps(1-5).

HotStart-IT SYBR Green One-Step qRT-PCR Master Mix Kit includes M-MLV RT, RNase Inhibitor, and a 2X Master Mix containing HotStart-IT Taq DNA Polymerase, MgCl2, Ultrapure nucleotides, and SYBR Green I in an optimized reaction buffer. HotStart-IT SYBR Green qPCR Master Mix uses a novel hot

start method developed at Affymetrix called primer sequestration. This novel hot start feature increases the specificity and sensitivity of SYBR-based qRT-PCR reactions by substantially reducing primer-dimer formation. With this method, the HotStart-IT protein binds and sequesters primers at lower tempera-tures making them unavailable for use by Taq DNA Polymerase. Following reverse transcription and the subsequent heat denaturation step, the primer binding protein is inactivated and the primers are released.

This kit exhibits excellent sensitivity as it can detect fewer than 10 target copies, performs over a broad, linear dynamic range of 6 to 7 orders of magnitude, and is compatible with most real-time PCR instru-ments.

Based on Affymetrix primer sequestration technology�� Increases specificity and prevents primer-dimer formation�� No DNA template damage – no extensive heating step needed to denature hotstart component

Shipping and storage:Shipped on dry ice. Store at -20°C. Mix all compo-nents well prior to use. Light sensitive components should be protected from excessive light exposure.

Components: 100 reaction kit 500 reaction kit Rctn. Vol. = 50 µl Rctn. Vol. = 50 µlHotStart-IT SYBR Green

qPCR Master Mix (2X) 2 x 1.25 ml 1 x 12.5 ml25 mM MgCl2 1 x 1 ml 5 x 1 mlROX Passive Reference Dye 1 x 100 μl 1 x 500 μlFluorescein Passive

Reference Dye 1 x 100 μl 1 x 500 μlM-MLV RT 1 x 40 μl 1 x 200 μlRNase Inhibitor (10 units/μl) 1 x 40 μl 1 x 200 μlRNase-Free Water,

DEPC-Treated 3 x 1 ml 1 x 15 ml







HotStart-IT SYBR Green One-Step qRT-PCR Master Mix Kit


Description:HotStart-IT Probe One-Step qRT-PCR Master Mix Kit provides optimal performance and maximum convenience for real-time, quantitative analysis of RNA templates in a single reaction format. The RT-PCR process converts and amplifies single-stranded RNA template yielding double-stranded DNA product. One-Step RT-PCR uses gene specific primers, designed to match RNA/cDNA targets, in a single-tube/plate, one-step reaction. This approach offers tremendous convenience when applied to analysis of single targets from multiple RNA samples. This minimizes the possibility of introducing con-taminants into reactions between the RT and PCR steps, since both steps are carried out sequentially without opening the reaction tubes/plates between the steps(1-5).

HotStart-IT Probe One-Step qRT-PCR Master Mix Kit includes M-MLV RT, RNase Inhibitor, and a 2X Mas-ter Mix containing HotStart-IT Taq DNA Polymerase, MgCl2, and Ultrapure nucleotides in an optimized reaction buffer for use with fluorescent probes. HotStart-IT Probe qPCR Master Mix uses a novel hot start method developed at Affymetrix called primer

sequestration (see PN 71195). With this method, the HotStart-IT protein binds and sequesters primers at lower temperatures making them unavailable for use by Taq DNA Polymerase. Following reverse transcription and the subsequent heat denaturation step, the primer binding protein is inactivated and the primers are released.

This kit is formulated for use with fluorescent probes such as TaqMan Probes, Molecular Beacons, and others(6-7). Since fluorescent probes are designed to hybridize to the target of interest, detection specificity is greatly increased relative to non-specific dsDNA binding dyes such as SYBR Green I. The Taq DNA Polymerase used in this master mix has the 5' to 3' exonuclease activity necessary for efficient removal of the 5'-fluorophore from the 3'-quencher in TaqMan probes.

Based on Affymetrix primer sequestration technology�� Increases specificity and prevents primer-dimer formation�� No DNA template damage – no extensive heating step needed to denature hotstart component

Shipping and storage:Shipped on dry ice. Store at -20°C. Mix all compo-nents well prior to use. Light sensitive components should be protected from excessive light exposure.

Components: 100 reaction kit 500 reaction kit Rctn. Vol. = 50 µl Rctn. Vol. = 50 µl

HotStart-IT Probe qPCR Master Mix (2X) 2 x 1.25 ml 1 x 12.5 ml

25 mM MgCl2 1 x 1 ml 5 x 1 mlROX Passive Reference Dye 1 x 100 μl 1 x 500 μlM-MLV RT 1 x 40 μl 1 x 200 μlRNase Inhibitor (10 units/μl) 1 x 40 μl 1 x 200 μlRNase-Free Water,

DEPC-Treated 3 x 1 ml 1 x 15 ml







6. Livak, K. J., Flood, S. J., Marmaro, J., Giusti, W., and Deetz, K. (1995) PCR Methods Appl. 4, 357-362.

7. Tyagi, S., and Kramer, F. R. (1996) Nat. Biotechnol. 14, 303-308.

HotStart-IT Probe One-Step qRT-PCR Master Mix Kit



78370 100 reactions

RT-PCR Master Mix provides maximum con-venience and optimal performance for highly sensitive and specific one-step RT-PCR reactions. This unique formulation combines all the reagents necessary for successful RT-PCR. Simply add RT-PCR Master Mix to RNA template, primers, and RNase-free water, and the reactions are ready to begin.

During reverse transcription-polymerase chain reac-tion (RT-PCR), reverse transcriptase converts RNA template to cDNA which is subsequently amplified by a thermostable DNA polymerase(1,2). One-step RT-PCR is a variation of RT-PCR in which the com-ponents necessary for both the RT and PCR steps are combined in a single-tube and the reactions are performed sequentially(2). This single-step, closed-tube approach simplifies the expression analyses of one or a few genes from multiple RNA samples and reduces the risk of contaminating samples. RT-PCR Master Mix further simplifies this technique by providing one-step RT-PCR in a pre-mixed format.

ConvenientRT-PCR Master Mix saves time and reduces potential contamination errors by eliminating several pipetting steps. Since the mix is pre-formulated and thoroughly QC tested, experimental variability is significantly reduced. This translates into greater reproducibility in demanding, high-throughput experiments. For a 50 µl reaction, simply add 25 µl of RT-PCR Master Mix to primers, RNA template, and RNase-free H2O.

Improve specificity and sensitivityAmplify RT-PCR products from less template, with lower background, and with little or no optimization. Targets may be routinely detected from just 100 fg of total RNA. For example, human β-actin is detected from 100 femtograms of total RNA, which represents about 1/100th of the total RNA of a single human cell (Fig. 1).

Optimal formulationAn enhanced buffer allows for RT reaction tempera-tures up to 50°C, which can improve detection of more difficult targets. This is because higher RT tem-peratures reduce non-specific priming and facilitate melting of RNA secondary structure(3). In addition, the mix has the wild-type, RNase H-plus form of M-MLV Reverse Transcriptase which has been shown to improve the sensitivity for certain targets(4). The RNase H activity degrades the RNA part of the cDNA/RNA hybrid following reverse transcription which may prevent its inhibition during subsequent PCR steps. RT-PCR Master Mix has been specifically designed to detect targets whose sizes are generally less than 1.0 kb. For targets greater than 1.0 kb and for high fidelity, use FideliTaq RT-PCR Master Mix (PN 71185).

StableRT-PCR Master Mix withstands repeated freeze-thaw cycles with no observed decrease in performance (Fig. 2).

Functional test:Tested by amplifying a 459 bp β-actin target from 10 pg of human placental total RNA.

RT-PCR Master Mix Formulation (2X):Includes M-MLV Reverse Transcriptase, Taq DNA Polymerase, recombinant RNase Inhibitor, nucleo-tides, and magnesium in a novel RT-PCR buffer. Magnesium concentration is 3 mM in the 2X RT-PCR Master Mix.

Components:RT-PCR Master Mix is supplied in kit form with the following components sufficient for 100 reactions in a 50 µl reaction volume:

4 x 625 µl RT-PCR Master Mix (2X)3 x 1 ml RNase-free Water1 x 1 ml 25 mM MgCl2 Solution


References:1. Sambrook, J. and Russell, D. W. (2001) “Molecular


2. Sellner, L. N., Coelen, R. J., and Mackenzie, J. S. (1992) Nucl. Acids Res. 20, 1487-1490.

3. Malboeuf, C. M., Isaacs, S. J., Tran, N. H., and Kim, B. (2001) BioTechniques 30, 1074-1084.

4. Polumuri, S. K., Ruknudin, A., and Schulze, D. H. (2002) BioTechniques 32, 1224-1225.

RT-PCR Master Mix (2X)

-RNA

1 kb –

500 bp –

200 bp –

M -RT 10 pg1 pg 100 fg

– β-actin (459 bp)

Total RNA

M -RNA -RT pre post pre post pre post pre post

10 ng 1 ng 100 pg 10 pg Total RNA

1 kb –500 bp –

200 bp –

– numb (455 bp)


1 ng 100 pg 10 pg 1 pg Total RNA

1 kb –500 bp –

200 bp –– β-actin (459 bp)

Fig. 1. Sensitivity of the RT-PCR Master Mix. A 459 bp frag-ment of the human β-actin gene was RT-PCR amplified from the indicated amounts of human placental total RNA. RT temperature was 50°C and primers were at 0.8 µM. The -RNA control included no RNA in the sample and the -RT control included 100 ng of total RNA but used the Taq PCR Master Mix (PN 71162, without reverse transcriptase) to test for any contaminating genomic DNA in the RNA sample. “M” is the DNA marker lane. Since the typical human cell contains about 10 picograms of total RNA, this target’s detection at 100 femtograms represents about 1/100th of the total RNA of a typical human cell.

Fig. 2. Stability of the RT-PCR Master Mix. Targets were RT-PCR amplified from the indicated amounts of human placental total RNA. RT temperature was 50°C and primers were at 0.8 µM. The -RNA control included no RNA in the sample and the -RT control includ-ed 100 ng of total RNA but used the Taq PCR Master Mix (PN 71162, without reverse tran-scriptase) to test for any contaminating genomic DNA in the RNA sample. “Pre” indicates lanes in which the Master Mix was not freeze-thawed and “Post” indicates lanes in which the Master Mix was freeze-thawed 15 times, alternating between dry ice and a room tem-perature water bath. “M” is the DNA marker lane.

PCR Tools | Reverse Transcription RT-PCR—One-Step Master Mix


71185 100 reactions

FideliTaq RT-PCR Master Mix provides maximum convenience and optimal performance for highly sensitive, specific, and accurate one-step RT-PCR reactions. This unique formulation combines all the reagents necessary for successful, high-fidelity RT-PCR. Simply add FideliTaq RT-PCR Master Mix to RNA template, primers, and RNase-free water, and the reactions are ready to begin.

During reverse transcription-polymerase chain reaction (RT-PCR), reverse transcriptase converts RNA template to cDNA which is subsequently amplified by a thermostable DNA polymerase(1,2). One-step RT-PCR is a variation of RT-PCR in which the components necessary for both the RT and PCR steps are combined in a single tube and the reactions are performed sequentially(2). This single-step, closed-tube approach simplifies the expression analyses of one or a few genes from multiple RNA samples and reduces the risk of contaminating samples. FideliTaq RT-PCR Master Mix further simplifies this technique by providing high-fidelity, one-step RT-PCR in a pre-mixed format.

ConvenientFideliTaq RT-PCR Master Mix saves time and reduces potential contamination errors by eliminating several pipetting steps. Since the mix is pre-formulated and thoroughly QC tested, experimental variability is significantly reduced. This translates into greater reproducibility in demanding, high-throughput experiments. For a 50 µl reaction, simply add 25 µl of FideliTaq RT-PCR Master Mix to primers, RNA template, and RNase-free H2O.

High fidelity and longer RT-PCR productsBy incorporating FideliTaq Polymerase into the mix, amplification fidelity is increased up to 6 times over Taq Polymerase alone, which is ideal for cloning applications(3-6). FideliTaq RT-PCR Master Mix gener-ates products with both blunt-ends and those with non-template added adenine on the 3' end. The ratio of blunt-ends to non-template, adenylated-ends varies from primer to primer and base identity at the 3' end(7). Thus, RT-PCR products may be cloned into both blunt-end and TA vectors, although TA vectors may yield more transformants. In addition, longer product sizes can be generated up to about 6 kb (Fig. 1).

Improve specificity and sensitivityAmplify RT-PCR products from less template, with lower background, and with little or no optimization. Targets may be routinely detected from just 100 fg of total RNA. For example, human β-actin is detected from 100 femtograms of total RNA, which represents about 1/100th of the total RNA of a single human cell.

Optimal formulationAn enhanced buffer allows for RT reaction tem-peratures up to 50°C, which can improve detection of more difficult targets. This is because higher RT temperatures reduce non-specific priming and facilitate melting of RNA secondary structure(8). In addition, the mix has the wild-type, RNase H-plus form of M-MLV Reverse Transcriptase which has been shown to improve the sensitivity for certain targets(9). The RNase H activity degrades the RNA part of the cDNA/RNA hybrid following reverse transcription which may prevent its inhibition during subsequent PCR steps.

Stable performanceThe FideliTaq RT-PCR Master Mix withstands repeat-ed freeze-thaw cycles with no observed decrease in performance (Fig. 1).

Functional test:Tested by amplifying a 459 bp β-actin target from 10 pg and a 1.5 kb β-actin target from 100 pg of human placental total RNA, plus a 5.6 kb Clathrin target from 10 ng of pooled human total RNA.

FideliTaq RT-PCR Master Mix Formulation (2X):Includes M-MLV Reverse Transcriptase, FideliTaq DNA Polymerase, recombinant RNase Inhibitor, nucleotides, and magnesium in a novel RT-PCR buffer. Magnesium concentration is 3 mM in the 2X FideliTaq RT-PCR Master Mix.

Components:FideliTaq RT-PCR Master Mix is supplied in kit form with the following components sufficient for 100 reactions in a 50 µl reaction volume:

4 x 625 µl FideliTaq RT-PCR Master Mix (2X)3 x 1 ml RNase-free Water1 x 1 ml 25 mM MgCl2 Solution


References:1. Sambrook, J. and Russell, D. W. (2001) “Molecular



3. Barnes, W. M. (1994) Proc. Natl. Acad. Sci. USA 91, 2216-2220.

4. Cheng, S., Fockler, C., Barnes, W. M., and Higuchi, R. (1994) Proc. Natl. Acad. Sci. USA 91, 5695-5699.




8. Malboeuf, C. M., Isaacs, S. J., Tran, N. H., and Kim, B. (2001) BioTechniques 30, 1074-1084.

9. Polumuri, S. K., Ruknudin, A., and Schulze, D. H. (2002) BioTechniques 32, 1224-1225.

FideliTaq RT-PCR Master Mix (2X)


10 ng 1 ng 100 pg 10 pg Total RNA

3 kb –

1.65 kb –1 kb –

– β-actin (1.5 kb)

M -RNA -RT pre post pre post pre post

100 ng 10 ng 1 ng Total RNA

12 kb –

3 kb –1.65 kb –

– Clathrin (5.6 kb)

Fig. 1. Stability and product length associated with the FideliTaq RT-PCR Master Mix. Targets were RT-PCR amplified from the indicated amounts of human total RNA. RT temperature was 50°C and primers were at 0.8 µM. The -RNA control included no RNA in the sample and the -RT control included 100 ng of total RNA but used the Taq PCR Master Mix (PN 71162, without reverse transcriptase) to test for any contaminating genomic DNA in the RNA sample. “Pre” indicates lanes in which the Master Mix was not freeze-thawed and “Post” indicates lanes in which the Master Mix was freeze-thawed 15 times alternating between dry ice and a room temperature water bath. “M” is the DNA marker lane. Human placental RNA was used for actin, and pooled RNA from several human cell-lines was used for Clathrin. The mix withstands repeated freeze-thaw cycles (up to 15) with no effect on product yield and is able to generate targets at least up to 5.6 kb.

PCR Tools | Reverse Transcription RT-PCR—One-Step Master Mix


78350 50 reactions

The One-Step RT-PCR Kit is designed for simple RT-PCR in a one-tube format.�� Streamlines and optimizes the RT and PCR steps�� Provides a starting point for analysis of new RNA targets

Reverse transcription-polymerase chain reaction, or RT-PCR, is a method for converting and amplifying a single-stranded RNA template to yield abundant double-stranded complementary DNA (cDNA) product(1,2). In the RT step, the RT enzyme reverse transcribes an RNA template, yielding single-stranded cDNA. In the PCR step, a thermostable DNA polymerase amplifies the single-stranded cDNA to yield double-stranded cDNA product. One step RT-PCR is a variation on RT-PCR in which all reaction components are mixed in one tube prior to starting the reactions(2). This approach offers simplicity and convenience and minimizes the possibility for contamination.

Complete One-Step RT-PCR KitThe One-Step RT-PCR Kit comes complete and ready-to-use for RT-PCR. The kit can be used with diverse RNA samples and custom primers. The kit uses M-MLV Reverse Transcriptase(3) and Taq DNA Polymerase(4), premixed together at concentrations optimized to balance sensitivity and specificity. An optimized reaction buffer, RNase Inhibitor, Ultrapure dNTPs, supplemental magnesium chloride, and RNase-free water are also included.

Highly sensitive, highly specificThe kit can be used for detection of diverse RNA targets (Fig. 1). Moderately or weakly expressed targets can be detected in 1 ng to 1 µg total RNA or 100 pg to 100 ng polyA RNA. Highly expressed tar-gets can be detected in even lower amounts of RNA (Fig. 2). Standard reaction conditions are sufficient for amplification of most targets. A few simple proto-col adjustments, such as optimization of the amount of primers or addition of supplements for amplifying G+C rich targets, enable specific amplification of other targets(5) (Fig. 3).

Convenient, one-tube formatSetting up reactions is quick and simple, given that the RT and PCR steps are set up simultaneously and then carried out sequentially in one tube. This format eliminates the need to set up RT and PCR indepen-dently, saving time and eliminating a potential point of contamination.

Rapid, reliable resultsThe One-Step RT-PCR Kit is ideal for qualitative analysis of expression of one or a few genes in multiple RNA samples, analysis of specific RNA splice variants, and streamlining the optimization of RT and PCR steps simultaneously. The kit is also particularly good as a starting point for analysis of new RNA targets, given that many targets can be amplified successfully without need for optimization.

Kit components:RT-PCR Enzyme Mix5X RT-PCR Reaction Buffer (includes MgCl2)Ultrapure PCR Nucleotide Mix (10 mM each dATP,

dCTP, dGTP, dTTP)RNase Inhibitor, Recombinant (4 units/µl)25 mM Magnesium ChlorideRNase-Free Water


References:1. Sambrook, J. and Russel, D. W. (2001) “Molecular





5. Tech Tip 206, Simple Approaches for Optimization of RT-PCR, USB Corporation, Cleveland, Ohio.

One-Step RT-PCR Kit

75780 50 reactions (20 µl)See page 39

First-Strand cDNA Synthesis Kit for Real-Time PCR

Fig. 1. Amplification of diverse RNA targets by one-step RT-PCR. Target (source): β-actin (100 ng total RNA, human liver), Numb (100 ng total RNA, human liver), Ubiquitin (1 µg total RNA, Arabidopsis leaf), and Terminal Deoxynucleotidyl Transferase (TdT) (100 ng polyA RNA, calf thymus). Gene specific primers were designed to generate products of particular sizes. M: Marker; Lanes 1-3: Actin, 0.5 kb, 1.0 kb, 1.5 kb; Lanes 4-5: Numb, 0.5 kb, 1.0 kb; Lane 6: Ubiquitin, 1.5 kb; Lane 7: TdT, 1.5 kb.

0.5 kb –

1 kb –

2 kb –

Fig. 2. Highly sensitive detection of β-actin target from human liver total RNA and polyA RNA, by one-step RT-PCR. Primers were used at 0.8 µM, a rela-tively high concentration, in order to achieve high sensitivity. Total RNA: 1 µg to 1 pg, 10-fold dilutions shown in Lanes 1-7. Poly A RNA: 100 ng to 100 fg, 10-fold dilutions shown in Lanes 1-7.

0.2 kb –

0.5 kb –

1.0 kb –

TotalRNA

PolyARNA

Fig. 3. Flexibility for optimization. For targets with high G+C contents, such as 0.34 kb Notch3 (G+C: 77%), adding supplements and/or increasing the temperature of the reverse transcription step, may improve specificity. Compare results for standard reac-tion conditions versus reaction with betaine supple-ment and elevated temperature.

0.2 kb –

0.5 kb –

1.5 M betaine:Temp (°C):

PCR Tools | Reverse Transcription RT-PCR—Kits


78355 50 RT rctns and 100 PCR rctns

The Two-Step RT-PCR Kit is designed for flexible RT-PCR in one- or two-tube formats.

�� Optimization of PCR independent of RT�� Analysis of the expression of multiple genes in individual RNA samples with oligo dT primers

Reverse transcription-polymerase chain reaction, or RT-PCR, is a method for converting and amplifying a single-stranded RNA template to yield abundant double-stranded complementary DNA (cDNA) product(1,2). In the RT step, the RT enzyme reverse transcribes an RNA template, yielding single-stranded cDNA. In the PCR step, a thermostable DNA polymerase amplifies the single-stranded cDNA to yield double-stranded cDNA product. RT-PCR may be carried out in several ways. One approach, often termed two-step RT-PCR, involves carrying out the RT (or cDNA synthesis) step in one tube and the PCR step in another tube(1) with gene specific primers, for detection of a single target from a single RNA sample. Another approach involves using one RT reaction, primed with oligo dT, for detection of multiple targets from the same RNA sample. A third approach, often termed one-step RT-PCR, involves setting up the RT and PCR simultaneously with gene specific primers and carrying out the reactions sequentially in one tube(2). This is particularly useful for detection of one or a few targets in multiple RNA samples. The Two-Step RT-PCR Kit has been designed for use in all of these types of RT-PCR.

All in one RT-PCR kitThe Two-Step RT-PCR Kit comes complete and ready-to-use for diverse RT-PCR applications. The kit can be used with RNA samples and custom primers. The kit uses M-MLV Reverse Transcriptase(3) and Taq DNA Polymerase(4), provided in individual tubes at concentrations optimized to balance sensitivity and specificity in two-step RT-PCR. Optimized RT and PCR reaction buffers, RNase Inhibitor, Ultrapure dNTPs, supplemental magnesium chloride, and RNase-free water are also included, allowing flex-ibility in setting up RT and PCR individually. A simple enzyme dilution step also allows use of the kit for one-step RT-PCR.

Highly sensitive, highly specificThe kit can be used for detection of diverse RNA targets based on generation of long cDNAs (to at least 5.6 kb) followed by amplification of short (~0.2 to 1.5 kb) PCR products (Fig. 1). Moderately and weakly expressed targets can generally be detected in 1 ng to 1 µg total RNA or 100 pg to 100 ng polyA RNA. Highly expressed targets can be detected in even lower amounts of RNA. Standard reaction conditions are sufficient for specific amplification of most targets. A few simple protocol adjustments, such as optimization of the amount of primers or addition of supplements for amplifying G+C rich targets, enable specific amplification for others(5).

Flexible formatA wide range of RT-PCR experiments are accessible due to the all-in-one nature of this kit. One set of reagents can be applied easily in multiple types of RT-PCR, allowing great flexibility in planning, carry-ing out, and optimizing experiments.

One kit, many applicationsThe Two-Step RT-PCR Kit is well suited for diverse RT-PCR applications. The two-step/gene specific primer format is excellent for qualitative analysis of gene expression, highly detailed analysis of RNA splice variants, and optimization of PCR indepen-dent of RT. The two-step/oligo dT primer format is excellent for analysis of the expression of multiple genes in individual RNA samples. And the one-step format is excellent for rapid analysis of single genes in multiple RNA samples. Depending on the scale of experiments and the size of PCR products, use of the One-Step RT-PCR Kit (PN 78350) coupled with the Taq PCR Master Mix or Taq PCR Kit may be more efficient. The Two-Step RT-PCR Kit (PN 78355) is one kit with many applications.

Kit components:M-MLV Reverse TranscriptaseTaq DNA PolymeraseRT Reaction Buffer (5X) (includes MgCl2)PCR Reaction Buffer (10X) (includes MgCl2)PCR Nucleotide Mix, Ultrapure: 10 mM each dATP,

dCTP, dGTP, dTTPRNase Inhibitor, Recombinant (4 units/µl)Magnesium Chloride (25 mM)RNase-Free (DEPC-treated) Water


References:1. Sambrook, J. and Russel, D. W. (2001) “Molecular





5. Tech Tip 206, Simple Approaches for Optimization of RT-PCR, USB Corporation, Cleveland, Ohio.

Related products

RT-PCR Master Mix (2X)78370 100 reactions

FideliTaq RT-PCR Master Mix (2X)71185 100 reactions

Two-Step RT-PCR Kit

Fig. 1. Amplification of diverse RNA targets by two-step RT-PCR. Target (source): β-actin (human liver), Numb (human liver), Ubiquitin (Arabidopsis leaf), and Terminal Deoxynucleotidyl Transferase (TdT) (calf thymus). RT was carried out on 1 µg total RNA with priming by oligo dT, and PCR was conducted on 10-1 dilution of RT reaction. Gene specific primers were designed to generate prod-ucts of particular sizes. M: Marker; Lanes 1-3: β-actin, 0.5 kb, 1.0 kb, 1.5 kb; Lanes 4-5: Numb 0.5 kb, 1.0 kb; Lane 6: Ubiquitin, 1.5 kb; Lane 7: TdT, 1.5 kb; Lane 8: Clathrin, 5.6 kb.

PCR Tools | Reverse Transcription RT-PCR—Kits


78250 20 reactions78200 100 reactions78201 500 reactions78202 2,000 reactions78205 5,000 reactionsCustom formulations by request

�� Conserves PCR samples – 100% recovery of both short and long PCR products�� One tube/one-step PCR cleanup – Add ExoSAP-IT reagent directly to PCR product.�� Eliminates spin columns – Decreases time and expense while increasing yield�� Removes contaminating primers and dNTPs – No interference in downstream applications�� Scalable – Economical for high-throughput purification�� Simple processing – Robotic-friendly; Replaces beads, filtrations, and plates�� Generates less waste than columns

ExoSAP-IT reagent is designed for simple, quick PCR cleanup for downstream applications, such as DNA sequencing or Single Nucleotide Polymorphism (SNP) analysis. When PCR amplification is complete, any unused dNTPs and primers remaining in the PCR product mixture will interfere with these methods. ExoSAP-IT PCR Cleanup removes these contaminants.

ExoSAP-IT reagent is added directly to the PCR product and incubated at 37°C for 15 minutes. After PCR treatment, it is inactivated simply by heating to 80°C for 15 minutes (Fig. 1).

ExoSAP-IT single-step PCR cleanup utilizes two hydrolytic enzymes, Exonuclease I and Shrimp Alkaline Phosphatase (SAP), together in a specially formulated buffer, to remove unwanted dNTPs and primers from PCR products. Exonuclease I removes residual single-stranded primers and any extraneous single-stranded DNA produced in the PCR. SAP removes the remaining dNTPs from the PCR mixture.

Rapid PCR product cleanup protocolThe ExoSAP-IT method requires only one pipet-ting step and two incubations. Just add ExoSAP-IT reagent to the PCR product and within 30 minutes sequencing or SNP analysis can be performed.

Simple: single-stepThe method is designed to require a minimum of “hands-on” time. Enzymatic removal of excess primers and unincorporated nucleotides occurs in one easy step by using ExoSAP-IT reagent in a single tube or microtiter well. Only simple pipette transfers are required, therefore, many samples can be pro-cessed at once, either manually or with robotics.

No sample lossUse of ExoSAP-IT reagent eliminates all gel or column purifications, sedimentations, filtrations, beads, and/or magnetic separations(1). There is 100% recovery of both short and long PCR products with ExoSAP-IT (Fig. 2).

Achieve high quality data from PCR productsExoSAP-IT reagent may be used as an effective cleanup method prior to fluorescent or radioactive DNA sequencing (Fig. 3), SNP analysis, or any other application requiring a PCR product free of excess nucleotides and primers.


References:1. Dugan, K. A., Lawrence, H. S., Hares, D. R.,

Fisher, C. L. and Budowle B. (2002) J. Forensic Sci 47, 811-818.

2. Hanke, M. and Wink, M. (1994) BioTechniques 17, 858-860.

3. Mu, J., Duan, J., Makova, K., Joy, D., Huynh, C., Branch, O., Li, W. and Su, X. (2002) Nature 418, 323-326.

4. Silva, Jr., W. A., Costa, M. C. R., Valente, V., De Freitas Sousa, J., Pacó-Larson, M. L., Espreafico, E. M., Camargo, S. S., Monteiro, E., De Jesus, A., Holanda, M. A., Zago, M. A., Simpson, A. J. G. and Neto, E. D. (2001) BioTechniques 30, 537-542.

5. Werle, E., Scneider C., Renner, M., Völker, M. and Fiehn, W. (1994) Nucl. Acids Res. 22, 4354-4355.

ExoSAP-IT PCR Product Cleanup

Fig. 2. ExoSAP-IT treatment of PCR products with no sample loss. Single-copy targets were amplified from human genomic DNA. HES-1 (125 bp), numb (455 bp), NRAGE (1.55 kb), and numb (4.6 kb) were loaded on a 1.5% agarose gel before (pre) and after (post) ExoSAP-IT treatment. M is the DNA marker lane. Note that a variety of PCR product sizes can be treated with ExoSAP-IT, even a 125 bp fragment, with no sample loss.

HES-1 numb NRAGE numb M pre post pre post pre post pre post

12 kb –

2 kb –

1 kb –

100 bp –

PCR Mixture Post-Amplification

PCR Product

+

Nucleosides

+

Inorganic Phosphate (Pi)

Add ExoSAP-IT®

Excess Primers

Excess dNTPs

37°C, 15 min for treatment80°C, 15 min to inactivate

Fig. 1. Summary of ExoSAP-IT PCR product treatment.

Fig. 3. Fluorescent sequencing results of a 100 bp pUC18 PCR fragment sequenced with a -20 Fwd primer using fluo-rescent sequencing reagents. PCR cleanup performed with: (a) ExoSAP-IT; (b) a column designed for PCR cleanup. Base miscalls in (b) are due to inherently low yields of short PCR products when using columns.

a

b

PCR Tools | PCR Purification

Related product

HT ExoSAP-IT High-Throughput PCR Product Cleanup78395 480 rctns x 8-tube strip

5,760 rctns x 1 plate (12 x 8-tube strips)

23,040 rctns - 4 plates1,000 rctns (2 ml)

5,000 rctns (10 ml)


78395 480 rctns x 8-tube strip5,760 rctns x 1 plate

(12 x 8-tube strips)23,040 rctns - 4 plates

1,000 rctns (2 ml)5,000 rctns (10 ml)

�� One-tube PCR cleanup – Add HT ExoSAP-IT reagent directly to PCR product.�� Exceptional accuracy – Achieve high quality data even with long read lengths (Fig. 1)�� 100% sample recovery – No loss of PCR products regardless of the fragment size (Fig. 2)�� Simple, single-step – Replace multiple steps and wasted time with bead or column use�� High-throughput processing – Even faster time to results with a low viscosity formulation, allowing robotic pipetting�� Removes excess primers and dNTPs – Does not interfere with downstream applications • sequencing • SNP analysis • single base extension • fragment analysis • in vitro transcription�� Scalable – Treat reaction volumes from 5 µl to 5 L�� Convenient packaging – Available in 8-tube strips and 12 x 8-tube strips in a 96-well plate �� Stable at +25°C for 8 hours – Retains full functional activity and at 4°C is stable for one week (Fig. 3)

HT ExoSAP-IT High-Throughput PCR Product Cleanup is an alternative formulation of ExoSAP-IT reagent specifically designed for the unique require-ments of high-throughput, automated platform and multi-channel pipettes. HT ExoSAP-IT reagent has both a longer lifetime at higher temperatures and a decreased viscosity for robotic pipetting. Like ExoSAP-IT PCR Product Cleanup (PN 78200), HT ExoSAP-IT reagent is a mixture of Exonuclease I and Shrimp Alkaline Phosphatase (SAP) which

removes excess primers and dNTPs following a PCR reaction in a single incubation. HT ExoSAP-IT reagent possesses greater thermal stability and lower viscosity which provides greater ease-of-use in automated 96- and 384-well formats.

HT ExoSAP-IT reagent is added directly to the PCR product and incubated at 37°C for 15 minutes. After PCR treatment, it is inactivated simply by heating to 80°C for 15 minutes. Our high-throughput formula-tion is designed for robotic pipetting and multi-channel pipettes. HT ExoSAP-IT reagent has a lower viscosity than the standard mix and is available in an 8-tube strip format for ease of use.

HT ExoSAP-IT reagent utilizes two hydrolytic enzymes, Exonuclease I and SAP, together in a specially formulated buffer, to remove unwanted dNTPs and primers from PCR products. Exonuclease I removes residual single-stranded primers and any extraneous single-stranded DNA produced in the PCR. SAP removes the remaining dNTPs from the PCR mixture.

Simple: Single-stepHT ExoSAP-IT reagent requires only one pipetting step and two incubations. Just add HT ExoSAP-IT reagent to the PCR product and within 30 minutes sequencing, SNP analysis, single base extension, or fragment analysis can be performed.

Enzymatic removal of excess primers and unincorpo-rated nucleotides occurs in one easy step by using HT ExoSAP-IT reagent in a single-tube, 8-tube strip, or 12 strips in a 96-well low skirted PCR plate. Our new formatting makes this an ideal product for use with robotics.

Exceptional accuracy with HT ExoSAP-IT Achieve high data quality and sequencing accuracy with HT ExoSAP-IT reagent, even with long read lengths. Sequence reads of PCR products treated with HT ExoSAP-IT reagent were on average 50+ bases longer than samples treated with competitor products. Using human genomic DNA, a 1,007 bp fragment was sequenced with no miscalls (Fig. 1). The size limitations associated with alternative PCR cleanup methods are not a factor with HT ExoSAP-IT reagent. Phred 20 values over the entire length of a 970 bp sequence was 822 ± 9 with HT ExoSAP-IT-treated samples and only 776 ± 83 with samples treated with Agencourt® AMPure® XP (see Table 1).

No sample lossUse of HT ExoSAP-IT eliminates all gel or column purifications, sedimentations, filtrations, beads, and/or magnetic separations(1). There is 100% recovery of both short and long PCR products with HT ExoSAP-IT High-Throughput PCR Product Cleanup (Fig. 2).

HT ExoSAP-IT High-Throughput PCR Product Cleanup

HT ExoSAP-IT

AMPure XP

3 miscalls

Fig. 2. Greater recovery of PCR product with HT ExoSAP-IT reagent. Equivalent volumes of PCR product were visualized on an ethid-ium bromide agarose gel and the band volume determined using image analysis. HT-ExoSAP-IT reagent allowed recovery of 100% of the 86 and 103 bp PCR products but AMPure XP beads allowed only 8 and 14% recovery, respectively. Un – Untreated, HT – HT-ExoSAP-IT treated, XP – AMPure XP purified, 100 bp – Affymetrix 100 bp ladder, bands from 100 to 1,000 bp by 100 bp intervals (PN 76712).

86 bp 103 bp100 bp Un HT XP Un HT XP

545 bp 1,007 bp100 bp Un HT XP Un HT XP


Fig. 1. Sequencing of a 1,007 bp treated PCR product. A 1,007 bp fragment was amplified and treated with HT ExoSAP-IT reagent (above) or Agencourt AMPure XP beads (below) and sequenced. Pherograms revealed no miscalls with HT ExoSAP-IT but three miscalls with Agencourt AMPure XP beads at position 203, 204, and 220.

Phred score values of samples processed with HT ExoSAP-IT and AMPure XP Beads

Table 1. HT ExoSAP-IT reagent gives better sequencing results.

HT ExoSAP-ITAMPure XP beads

Number bases read (Average of 16 samples)

901 850

Total miscalls 0 25

Pass rate (%)* 100% 99.7%

Phred 20** 822 ± 9 776 ± 83

* Pass rate – Average Phred value greater than 20 bases between 100 bp and 300 bp.** Phred 20 values over the entire length of sequence (no trimming ~970 bp)


HT ExoSAP-IT-treated PCR product is stableHT ExoSAP-IT reagent is rigorously tested and subject to strict quality control. Fig. 3 demonstrates that no product degradation occurred after storage of HT ExoSAP-IT-treated PCR product for one week at 25°C.








Related products


VeriQuest Fast Probe qPCR Master Mix (2X)75680 100 reactions 500 reactions 1,000 reactions 2,500 reactions 5,000 reactions

VeriQuest Fast Probe qPCR Master Mix, No Reference Dye (2X)75685 100 reactions 500 reactions 1,000 reactions 2,500 reactions 5,000 reactions

VeriQuest Fast SYBR Green qPCR Master Mix (2X)75690 100 reactions 500 reactions 1,000 reactions 2,500 reactions 5,000 reactions

VeriQuest Fast SYBR Green qPCR Master Mix with Fluorescein (2X)75675 100 reactions 500 reactions 1,000 reactions 2,500 reactions 5,000 reactions

VeriQuest Probe qPCR Master Mix (2X)75650 40 reactions 200 reactions 400 reactions 1,000 reactions 2,000 reactions




VeriQuest SYBR Green qPCR Master Mix (2X)75600 40 reactions








FideliTaq DNA Polymerase71180 50 units

250 units1,000 units

5 x 250 units5,000 units

RubyTaq DNA Polymerase71190 50 units



Fig. 3. High stability of HT ExoSAP-IT-treated products. A 1,007 bp DNA fragment amplified from genomic DNA with HotStart FideliTaq Master Mix (PN 71156) was treated with HT ExoSAP-IT reagent and stored for seven days at: -20ºC (lane 1), 4ºC (lane 2), or 25ºC (lane 3). Samples were visualized on 1.5% agarose/TAE/ethid-ium bromide gel. Lane MW: DNA Ladder, 1 kb Plus (PN 76714).

1 kb -20°C 4°C 25°C

HT ExoSAP-IT High-Throughput PCR Product Cleanup continued...



70995 100 reactions70996 500 reactions70997 2,000 reactionsCustoms by request

The PCR Product Pre-Sequencing Kit uses a novel enzymatic cleanup method to pre-treat PCR products prior to sequencing without any subsequent purifica-tion or separation steps. The two hydrolytic enzymes used in this kit, Shrimp Alkaline Phosphatase (SAP) and Exonuclease I, effectively remove excess dNTPs and primers present in the final PCR product reac-tion mixture. The enzymes are conveniently added to an aliquot of the PCR product mixture, incubated at 37°C, and then inactivated at 80°C. The result is a cleaner PCR product, free from excessive nucleotides and primers, ready to be sequenced with standard sequencing reagents.

Achieve high sequence quality with PCR productsEnzymatic treatment to remove excess primers and nucleotides yields templates which can be easily sequenced. Problems with left-over PCR primers leading to background bands are virtually elimi-nated. The kit may be used as an effective cleanup method prior to any DNA sequencing protocol, including but not limited to the Sequenase Version 2.0 DNA Sequencing Kit (PN 70770) or the Thermo Sequenase Cycle Sequencing Kits (PNs 78500 and 79260).

Kit components:Shrimp Alkaline PhosphataseExonuclease I


References:1. Hanke, M. and Wink, M. (1994) BioTechniques

17, 858-860.2. Werle, E., Scneider C., Renner, M., Völker, M. and

Fiehn, W. (1994) Nucl. Acids Res. 22, 4354-4355.

Related products

Sequenase™ Version 2.0 DNASequencing Kit70770 100 reactions

Thermo Sequenase™ Cycle Sequencing Kit78500 100 reactions

PCR Product Pre-Sequencing Kit

78260 864 µl

SBE Cleanup Reagent is a specialized formulation of ExoSAP-IT PCR Product Cleanup (PN 78200), which is based on enzymatic degradation of unconsumed primers and dNTPs present in PCR products. SBE Cleanup Reagent has been designed for use with the Beckman Coulter BioSciences SNPware™ Core Reagent Kit (PN 101-04-300).

Single-stepThe SBE Cleanup Reagent is designed for simple, quick PCR cleanup. SBE Cleanup Reagent utilizes two hydrolytic enzymes, Exonuclease I and Shrimp Alkaline Phosphatase (SAP), together in a specially formulated buffer, to remove unwanted dNTPs and primers from PCR products. Exonuclease I removes residual single-stranded primers and any extraneous

single-stranded DNA produced in the PCR. SAP removes the remaining dNTPs from the PCR mixture. It is added directly to the PCR product. SBE Cleanup Reagent is active in the buffer used for PCR, so no buffer exchange is required.


SBE Cleanup Reagent

Binding capacity up to 15 µg78758 50 preps78759 250 prepsSee page 146

Designed for the purification of DNA from reactions or for buffer exchange

�� Spin columns based on familiar silica-membrane technology�� High recovery rates up to 90% with low elution volume (15 μl)

�� Ideal for cleanup of reaction products�� Binding capacity up to 15 μg�� For DNA fragments 50 – 10,000 bp

The PrepEase DNA Cleanup Kit is designed to purify DNA fragments from enzymatic reactions or for buffer exchange. Thus, contaminants such as salts, enzymes, nucleotides, labeling reagents, etc. can be removed from a DNA sample. This kit can also be used for purification of DNA from Chromatin Immunoprecipitation (ChIP) assays.

Each kit includes PrepEase Cleanup Columns, collection tubes and the necessary reagents for the purification of DNA fragments.

PrepEase DNA Cleanup Kits

Binding capacity up to 15 µg78756 50 preps78757 250 prepsSee page 146

Designed for purifying DNA fragments from agarose gels. Also suited for PCR product purification.

�� Spin columns based on familiar silica-membrane technology�� High recovery rates of up to 90% with low elution volume (15 µl)�� Ideal for purifying restriction-digested DNA in gels or PCR products

The PrepEase Gel Extraction Kit may be used to purify DNA from agarose gels or PCR reaction mixtures. In addition, the kit may be used for desali-nation, and the removal of enzymes, nucleotides or labeling reagents from sample DNA.

PrepEase Gel Extraction Kits



78761 10 x 96-well plates78762 50 x 96-well platesSee page 148

Designed for high-throughput purification of PCR products using 96-well plates.

�� For PCR products ≥ 150 bp�� Process 20 - 300 μl PCR volume per well�� Recover up to 95% depending on length of PCR product in as little as 25 μl

�� May be performed manually by vacuum, by centrifugation or by using an automated workstation�� Includes 96-well plates and buffer for PCR product purification

PrepEase PCR Purification 96-Well Plate Kits (Ultrafiltration)

75767 500 μl

Fluorescein Passive Reference Dye is specially formu-lated for use on Bio-Rad real-time PCR instruments such as the MyiQ, iCycler iQ, and iQ5. This inert dye may be added to quantitative, real-time PCR reac-tions containing SYBR Green I. Fluorescein allows the collection of Dynamic Well Factors from the experimental plate, which is the preferred method. This normalizes the well-to-well differences that may occur due to artifacts such as pipetting errors or instrument limitations.

Fluorescein Passive Reference Dye is composed of a 500 nM solution of fluorescein in 10 mM Tris-HCl (pH 8.6), 0.1 mM EDTA, and 0.01% Tween®-20. Generally, the optimum final fluorescein concentra-tion is 10 nM but a titration between 5-20 nM may be necessary depending on the particular instru-ment, the amount of SYBR in the reaction mix, and the desired background. See specific instrument instructions for further details on well factor collection. The following table shows current recommendations from Affymetrix.

Spectral Characteristics:Excitation λ Maximum: ≈ 495 nmEmission λ Maximum: ≈ 515 nm

Shipping and storage:Shipped on dry ice. Store at -20°C. Protect from light.

Fluorescein Passive Reference Dye

Fluorescein Final ConcentrationAmount per 50 μl reaction Final Concentration Dilution Factor

1.0 μl (0.5 - 2.0 μl) 10 nM (5 - 20 nM) 50X

Subtract the volume of the dye from the volume of water needed to prepare the PCR reaction.

75768 500 μl

ROX Passive Reference Dye is specially formulated for use on Applied Biosystems (ABI) and Stratagene real-time PCR instruments. This inert dye, whose fluorescence does not change during the reaction, may be added to quantitative, real-time PCR reac-tions to normalize the well-to-well differences that may occur due to artifacts such as pipetting errors or instrument limitations.

ROX Passive Reference Dye is composed of a 25 μM solution of 5-carboxy-X-rhodamine in 10 mM Tris-HCl (pH 8.6), 0.1 mM EDTA, and 0.01% Tween-20.

The following table shows current recommendations from Affymetrix.

Spectral Characteristics:Excitation λ Maximum: ≈ 575 nmEmission λ Maximum: ≈ 600 nm

Shipping and storage:Shipped on dry ice. Store at -20°C. Protect from light.

ROX Passive Reference Dye

ROX Final Concentration for Different Instruments Amount per Final ROX Dilution Instrument 50 μl reaction Concentration Factor

ABI 7000, 7300, 7700, 1.0 μl 500 nM 50X 7900HT, 7900HT Fast, (0.6 - 1.0 μl) (300 - 500 nM) StepOne™ and StepOne Plus™

ABI 7500 and ABI 7500 Fast; 0.1 μl 50 nM 500X Stratagene Mx3000™, (0.06 - 0.1 μl)* (30 - 50 nM) Mx3005P™, and Mx4000™

Subtract the volume of the dye from the volume of water needed to prepare the PCR reaction.*Note: ROX may be diluted 1:10 in water to aid in accurate pipetting.

PCR Tools | PCR Purification/Passive Reference Dyes

NucleotidesDeoxynucleotides

PCR Nucleotide Mixes

Dideoxynucleotides

Ribonucleotides

Labeled Nucleotides


USB Ultrapure Nucleotide Solutions

�� Functionally tested: All USB Ultrapure dNTPs are functionally tested in long PCR to generate a 20.7 kb fragment.�� Convenient packaging: dNTPs are available separately, as a set of 4, or as a PCR mix.�� Bulk sizes and custom blends available upon request.

Deoxynucleotides (dNTPs), Sodium Salt Aqueous Solutions, pH 7.5

Prod. No. Product Description [conc] Size

77102 dATP 100 mM 25 µmol (250 µl) 2'-Deoxyadenosine-5'-Triphosphate 100 µmol (1 ml) Ultrapure 500 µmol (5 ml)

77104 dCTP 100 mM 25 µmol (250 µl) 2'-Deoxycytidine-5'-Triphosphate 100 µmol (1 ml) Ultrapure 500 µmol (5 ml)

77106 dGTP 100 mM 25 µmol (250 µl) 2'-Deoxyguanosine-5'-Triphosphate 100 µmol (1 ml) Ultrapure 500 µmol (5 ml)

77108 dTTP 100 mM 25 µmol (250 µl) 2'-Deoxythymidine-5'-Triphosphate 100 µmol (1 ml) Ultrapure 500 µmol (5 ml)

77206 dUTP 100 mM 25 µmol (250 µl) 2'-Deoxyuridine-5'-Triphosphate 100 µmol (1 ml) Ultrapure 500 µmol (5 ml)

77100 dNTPs, Sets of Four (dA, dC, dG, dT) 100 mM 4 x 25 µmol (250 µl) 2'-Deoxynucleoside-5'-Triphosphates, Ultrapure 4 dNTPs per pack (pk) 1 pk77328 100 mM 4 x 100 µmol (1 ml) 4 dNTPs per pack (pk) 1 pk77128 100 mM 4 x 500 µmol (5 ml) 4 dNTPs per pack (pk) 1 pk

77212 PCR Nucleotide Mix, 10 mM Solution 10 mM 500 µl Ultrapure 10 mM 2 x 500 µl 10 mM each dATP, dCTP, dGTP, dTTP

77119 PCR Nucleotide Mix, 25 mM Solution 25 mM 500 µl Ultrapure 25 mM each dATP, dCTP, dGTP, dTTP

77330 PCR Nucleotide Mix with dUTP, 10 mM Solution 10 mM 500 µl Ultrapure 10 mM each dATP, dCTP, dGTP, dUTP

Deoxynucleotides (dNTPs), Lithium Salt Aqueous Solutions, pH 7.0


70065 7-deaza-dGTP 10 mM 2 µmol (200 µl) 7-deaza-2'-Deoxyguanosine-5'-Triphosphate Ultrapure

Nucleotides


Dideoxynucleotides (ddNTPs), Sodium Salt Aqueous Solutions, pH 7.5�� Functionally tested: USB ddNTPs are functionally tested to meet or exceed criteria for high quality sequencing.�� Bulk sizes and custom blends available upon request.


77110 ddATP 10 mM 0.5 µmol (50 µl) 2',3'-Dideoxyadenosine-5'-Triphosphate 1 µmol (100 µl) Ultrapure

77112 ddCTP 10 mM 0.5 µmol (50 µl) 2',3'-Dideoxycytidine-5'-Triphosphate 1 µmol (100 µl) Ultrapure

77116 ddTTP 10 mM 0.5 µmol (50 µl) 2',3'-Dideoxythymidine-5'-Triphosphate 1 µmol (100 µl) Ultrapure

77126 ddNTPs, Set of Four (ddA, ddC, ddG, ddT) 10 mM 4 x 0.5 µmol (50 µl) Dideoxynucleoside Triphosphates 4 ddNTPs per pack (pk) Ultrapure 1 pk77125 10 mM 4 x 1 µmol (100 µl) 4 ddNTPs per pack (pk) 1 pk77335 100 mM 4 x 4 µmol (40 µl) 4 ddNTPs per pack (pk) 1 pk

Ribonucleotides (NTPs), Sodium Salt Aqueous Solutions, pH 7.5


77241 ATP 100 mM 25 µmol (250 µl) Adenosine-5'-Triphosphate Ultrapure

77243 GTP 100 mM 25 µmol (250 µl) Guanosine-5'-Triphosphate Ultrapure

77245 NTPs, Set of Four (ATP, CTP, GTP, UTP) 100 mM 4 x 25 µmol (250 µl) Nucleoside Triphosphates 4 NTPs per pack (pk) Ultrapure 1 pk

USB Ultrapure Nucleotide Solutions

Nucleotides


Nucleotides | Nucleotide Labeling Reagents

Molecular λ max Absorbance at λ max Compound Weight (pH 7.0) 1 M solution (pH 7.0)

ATP 507.2 259 15,400

CTP 483.2 271 9,000

GTP 523.2 253 13,700

UTP 484.2 262 10,000

dATP 491.2 259 15,200

dCTP 467.2 271 9,300

dGTP 507.2 253 13,700

dTTP 482.2 267 9,600

Nucleotide Physical Properties

79015 250 nmol (10 mM) 1,000 nmol (10 mM)

DNA Labeling Reagent, DLR, is a proprietary biotin-labeled, abasic, deoxynucleotide analog designed for efficient 3’ end-labeling of DNA using Terminal Deoxynucleotidyl Transferase (TdT) after amplification reactions such as PCR. When used with streptadivin conjugated to a reporter, the 3’ biotin labeled DNA

fragment can be detected by enzyme activity, fluo-rescence, or by the binding of a secondary antibody. In addition, DLR will not base-pair with natural nucleotides, thus reducing non-specific binding and maximizing the signal-to-noise ratio. DLR is a com-ponent of the Affymetrix GeneChip DNA labeling reagent (PN 900542) and has been used extensively to successfully probe microarrays for decades.

C24H40Li4N5O16P3SFormula Weight: 807.35

Product specifications:Purity: >95%Concentration: 10 mMAppearance: Clear, colorless liquid

Storage: Store at -20°C, avoid freeze/thaw cycles

Biotin-11-dXTP Analog (DNA Labeling Reagent, DLR)

79010 250 nmol (10 mM) 1,000 nmol (10 mM)

RNA Labeling Reagent, RLR, is a proprietary biotin-labeled pseudo-uridine triphosphate designed for incorporation into an RNA strand during an in vitro transcription reaction. This creates an RNA target internally labeled with biotin that can be detected

using streptadivin conjugated to fluorophores, enzymes, or antibodies. RLR is designed for efficient incorporation by T7 RNA polymerase and has been extensively tested in the Affymetrix GeneChip IVT labeling kit (PN 900449). The reagent is optimized for use in high throughput applications and probing high density microarrays.

C30H44Li4N7O19P3SFormula Weight: 959.46

Product specifications:Purity: >95%Concentration: 10 mMAppearance: Clear, colorless liquid

Storage: Store at -20°C, avoid freeze/thaw cycles

Biotin-11-ψTP Analog (RNA Labeling Reagent, RLR)


Copyright 1983 from Molecular Biology of the Cell, by Alberts et al. Reproduced by permission of Garland Science/Taylor & Francis LLC.

Structures of Nucleotides and Nucleosides

Nucleotides


2'-Deoxyadenosine-5'-Triphosphate, Sodium Salt, 100 mM Solution (dATP)

ULTRAPUREC10H12N5O12P3Na4

Formula Weight: 579.11Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λ max: 259 nm ± 1 nmSpectral Ratios: A250/A260: 0.80 ± 0.03 A280/A260: 0.12 ± 0.02Functional Tests: Functionally tested in long PCR to

generate a 20.7 kb fragment.Storage: -20°C

77102 25 µmol (250 µl) 100 µmol (1 ml) 500 µmol (5 ml)

2'-Deoxycytidine-5'-Triphosphate, Sodium Salt, 100 mM Solution (dCTP)


Formula Weight: 555.08Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λ max (pH 2.0): 280 nm ± 1 nmSpectral Ratios (pH 2.0): A250/A260: 0.45 ± 0.03 A280/A260: 2.10 ± 0.15 A290/A260: 1.60 ± 0.10Functional Tests: Functionally tested in long PCR to



2'-Deoxyguanosine-5'-Triphosphate, Sodium Salt, 100 mM Solution (dGTP)


Formula Weight: 595.11Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λmax: 252 nm ± 1 nmSpectral Ratios (pH 7.5): A250/A260: 1.18 ± 0.04 A280/A260: 0.67 ± 0.03 A290/A260: 0.28 ± 0.03Functional Tests: Functionally tested in long PCR to



2'-Deoxythymidine-5'-Triphosphate, Sodium Salt, 100 mM Solution (dTTP)





2'-Deoxyuridine-5'-Triphosphate, Sodium Salt, 100 mM Solution (dUTP)


Formula Weight: 556.1Product specifications:Form: 100 mM aqueous solutionpH: 7.5 ± 0.2Assay (HPLC): ≥ 99%λmax: (pH 7.5) 262 nm ± 1 nmAbsorbance Ratios: A250/A260: 0.74 ± 0.03 A280/A260: 0.35 ± 0.03Functional Test: Functionally tested in PCRStorage: -20°C


2'-Deoxynucleoside-5'-Triphosphates, Sodium Salt, 100 mM Solutions, Sets of Four (dNTPs)

ULTRAPUREForm: 100 mM aqueous solutions, pH 7.5; dATP,

dCTP, dGTP, dTTPFunctional Tests: Functionally tested in long PCR to


4 x 25 µmol (250 µl) each dNTP per pack77100 1 pk4 x 100 µmol (1 ml) each dNTP per pack77328 1 pk4 x 500 µmol (5 ml) each dNTP per pack77128 1 pk

For powder forms of the above nucleotides and our complete selection of nucleotides, nucleosides, purines and pyrimidines, please see the Biochemicals section of this catalog.

Nucleotides | Deoxynucleotides (dNTPs)


PCR Nucleotide Mix, 10 mM SolutionULTRAPURE

Form: Premixed dNTPs at 10 mM each dATP, dCTP, dGTP, dTTP.

pH: 7.5Functionally tested in PCR with USB Taq DNA

Polymerase (PN 71160).Storage: -20°C

77212 500 µl 2 x 500 µl





77119 500 µl

PCR Nucleotide Mix with dUTP, 10 mM SolutionULTRAPURE

Form: Premixed dNTPs at 10 mM each dATP, dCTP, dGTP, dUTP



77330 500 µl

7-deaza-2'-Deoxyguanosine-5'-Triphosphate, Lithium Salt, 10 mM Solution (7-deaza-dGTP)

ULTRAPUREC11H15N4O13P3Li2Formula Weight: 518.10Product specifications:Form: 10 mM aqueous solutionAssay (HPLC): ≥ 95%pH: 7.0λmax (pH: 7.5): 259 nm ± 1 nmFunctional Tests: Meets or exceeds criteria for high

quality sequencing.Storage: -20°C

70065 2 µmol

2',3'-Dideoxyadenosine-5'-Triphosphate (ddATP)


Formula Weight: 563.11Product specifications:Assay (FPLC): ≥ 99%pH: 7.5 ± 0.2λmax: 259 nm ± 1 nmSpectral Ratios (pH 7.5): A250/A260: 0.80 ± 0.03 A280/A260: 0.12 ± 0.02Functional Test: Meets or exceeds criteria for high


10 mM aqueous solution, pH 7.577110 0.5 µmol (50 µl) 1 µmol (100 µl)

2',3'-Dideoxycytidine-5'-Triphosphate (ddCTP)


Formula Weight: 539.08Product specifications:Assay (FPLC): ≥ 99%pH: 7.5 ± 0.2λmax (pH 2.0): 280 nm ± 1 nmSpectral Ratios (pH 2.0): A250/A260: 0.45 ± 0.03

A280/A260: 2.20 ± 0.20A290/A260: 1.70 ± 0.20

Functional Test: Meets or exceeds criteria for high quality sequencing.

Storage: -20°C


2',3'-Dideoxythymidine-5'-Triphosphate (ddTTP)


Formula Weight: 554.10Product specifications:Assay (FPLC): ≥ 99%pH: 7.5 ± 0.2λmax (pH 7.5): 267 nm ± 1 nmSpectral Ratios (pH 7.5): A250/A260: 0.65 ± 0.03 A280/A260: 0.83 ± 0.03Functional Tests: Meets or exceeds criteria for high



2',3'-Dideoxynucleoside-5'-Triphosphates, 10 mM, Sets of Four (ddNTPs)

ULTRAPUREForm: 10 mM aqueous solutions, pH 7.5; ddATP,

ddCTP, ddGTP, ddTTPFunctional Tests: Meets or exceeds criteria for high


4 x 0.5 µmol (50 µl) each dNTP per pack77126 1 pk4 x 1.0 µmol (100 µl) each dNTP per pack77125 1 pk

2',3'-Dideoxynucleoside-5'-Triphosphates, 100 mM, Set of Four (ddNTPs)

ULTRAPUREForm: 100 mM aqueous solutions, pH 7.5; ddATP,

ddCTP, ddGTP, ddTTPFunctional Tests: Meets or exceeds criteria for high


4 x 4 µmol (40 µl) each ddNTP per pack77335 1 pk

Custom blends available upon request


Nucleotides | Deoxynucleotides (dNTPs) & Dideoxynucleotides (ddNTPs)


Adenosine-5'-Triphosphate, 100 mM Solution (ATP)


Formula Weight: 595.11Form: 100 mM aqueous solution, pH 7.5Storage: -20°C

77241 25 µmol (250 µl)

Guanosine-5'-Triphosphate, Sodium Salt, 100 mM Solution (GTP)



77243 25 µmol (250 µl)

Nucleoside-5'-Triphosphates (ATP, CTP, GTP, UTP), Set of Four, 100 mM Solutions (NTPs)

ULTRAPURE100 mM aqueous solutions, pH 7.5Storage: -20°C

4 x 25 µmol (250 µl) each NTP per pack.77245 1 pk

Related products – RNA analysis

Oligo(dT)12-18 Primer, MW ≅450077405 100 µl

Oligo p(dT)12-18, Sodium Salt19817 5 units 25 units 100 units

Poly(A) Polymerase, Yeast74225Y 10,000 units74225Z 25,000 units

T3 RNA Polymerase70051Y 1,000 units70051Z 5,000 units

T7 RNA PolymeraseStandard Concentration, 20 units/µl70047Y 6,000 unitsHigh Concentration, >200 units/µl70001Z 30,000 units

Nucleotides | Ribonucleotides (NTPs)


With over 40 years experience in the life science industry, we can assist you with custom reagent manufacturing, dispensing, labeling, and kitting of your critical reagent components.

Customized solutionsMost of our 3,000 catalog products can be tailored to meet your specifications. We provide: �� Custom concentrations�� Customer-specific quality control assays�� Private labeling (In-house printing or customer-supplied)�� Complete traceability of kit components

Tailored reagent fillingWe deliver flexibility in scale and process with:�� Customization with over 60 bottle/vial options�� Lot-to-lot consistency�� General Purpose Reagents (GPRs) on request�� Scalable capabilities for future growth

To learn more, visit usb.affymetrix.com/bulk.

USB offers custom and OEM solutions

Molecular Biology EnzymesBinding Proteins

Glycosylases

Kinases

Ligases

Nucleases

DNases

Endonucleases

Exonucleases

RNases

Phosphatases

Alkaline Phosphatases

Pyrophosphatase

Polymerases

DNA Polymerases

Reverse Transcriptases

RNA Polymerases

Transferase

RNase Inhibitors

Topoisomerase


Molecular Biology Enzymes | Table of Contents

Enzyme Applications 64

Binding Proteins RecAProtein 65Single-StrandedDNABindingProtein(SSB) 65T4Gene32Protein 66

GlycosylasesUracil-DNAGlycosylase,E. coli 67Uracil-DNAGlycosylase,Heat-Labile 67

KinasesOptiKinase™ 68T4PolynucleotideKinase(PNK) 69

LigasesE. coliDNALigase 70T4DNALigase 70T4RNALigase 71T4RNALigase2 71

Nucleases - DNasesDNaseI,Lyophilized 72DNaseI,Solution 72rDNaseI,RNase-Free 73ShrimpDNase,Recombinant 7410XDNaseIBuffer 74

Nucleases - EndonucleasesHumanApurinic/ApyrimidinicEndonuclease1

(APE1) 75T4EndonucleaseVII 75

Nucleases - ExonucleasesExonucleaseI 76ExonucleaseIII 77ExonucleaseVII 77T7Gene6Exonuclease 78

Nucleases - RNasesRNaseI 79RNaseI‘A’,Powder 79RNaseA,Lyophilized 79RNaseA,MBGrade,Solution 80RNaseH 80

Nucleases - Non-SpecificMicrococcalNuclease 81PhosphodiesteraseI 81

Phosphatases - Alkaline PhosphatasesCalfIntestinalAlkalinePhosphatase(CIAP) 82ShrimpAlkalinePhosphatase(SAP) 83SuperSAP™ShrimpAlkalinePhosphatase 84

Phosphatases - PyrophosphatasePyrophosphatase,Inorganic(Recombinant)

(rPPase) 85

Polymerases - DNA PolymerasesExonuclease-FreeKlenow 86Exonuclease-FreeKlenowTrisBuffer 86DNAPolymeraseI 87KlenowDNAPolymeraseI 87Sequenase™Version2.0DNAPolymerase 88T7DNAPolymerase 88TthDNAPolymerase 89

Polymerases - Reverse TranscriptasesAMVReverseTranscriptase 90M-MLVReverseTranscriptase 90

Polymerases - RNA PolymerasesE. coliRNAPolymeraseHoloenzyme 91Poly(A)Polymerase,Yeast 91T3RNAPolymerase 92T7RNAPolymerase 92

Polymerases - TransferaseTerminalDeoxynucleotidylTransferase,

Recombinant(rTdT) 93

RNase InhibitorsRNaseInhibitor(HumanPlacenta) 94RNaseInhibitor(Recombinant) 94

TopoisomeraseTopoisomeraseII,Alpha 95


Molecular Biology Enzymes | Enzyme Applications

Application Enzyme

Blunt-endgeneration byremovalof3'protrudingends................T4andT7DNAPolymerase,Klenowfragment*,Blunt-ITRepairKit* byfillingin3'recessedends ...................T4andT7DNAPolymerase;Klenowfragment,Blunt-ITRepairKit*cDNAsynthesis fromRNA .................................ReverseTranscriptase;AMVRTorM-MLVRT,TthDNAPolymerase* ofsecondstrand ............................Klenowfragment,Sequenase,TaqDNAPolymerase,FideliTaqDNAPolymerase*CloningofDNAfragments .........................ShrimpAlkalinePhosphatase(SAP)*;CIP;T4DNALigase;Klenowfragment;restrictionendonucleasesDegradationofdouble-&single-strandedDNA*.........DNaseI*Degradationofdouble-strandedDNA,leaving...........ShrimpDNase* single-strandedDNAintact*DegradationofDNA Exonucleases—double-stranded 5'→3'....................................T7Gene6andLambdaExonuclease 3'→5'....................................ExonucleaseIII 5'→3'and3'→5'...........................Bal 31Nuclease,ExonucleaseV* Exonucleases—single-stranded 3'→5'....................................ExonucleaseI 5'→3'and3'→5'...........................ExonucleaseVII*,ExonucleaseV*DegradationofRNA..............................RibonucleaseAand/orT1;micrococcalnucleaseDegradationofRNAinRNA:DNAduplexes.............RibonucleaseHEndonucleases—double-stranded non-specific................................DNaseI,Bal31,andmicrococcalnucleases specific ...................................RestrictionendonucleasesEndonucleases—single-stranded ....................S1,Bal 31andmungbeannucleasesDephosphorylation...............................ShrimpAlkalinePhosphatase(SAP)*,BAPandCIPDNAsequencing.................................ModifiedT7DNAPolymerase(Sequenase*);TaqPolymerase;Klenowfragment;T4PolynucleotideKinase;

ReverseTranscriptase;ThermoSequenase*Intronmapping..................................ExonucleaseVII;S1NucleaseLabelingDNAat5'ends...........................T4PolynucleotideKinase,OptiKinase*LabelingDNAat3'ends...........................ModifiedT7DNAPolymerase(Sequenase*);Klenowfragment;ReverseTranscriptase;TerminalTransferaseLabelingRNAat3'ends...........................Poly(A)Polymerase;T4RNALigaseLigationsofDNA.................................E. coli andT4DNALigase,Ligate-ITRapidLigationKit*LigationsofRNA.................................T4RNALigaseMappingmRNA5'ends...........................S1orMungBeanNuclease;ReverseTranscriptase;RibonucleaseAMethylationofDNA..............................MethylasesMultiplexPCR...................................MagniTaqMultiplexPCRMasterMix*Nesteddeletionconstruction........................Bal 31;ExonucleaseIII;S1orMungBeanNucleaseNicktranslation..................................DNaseI;E. coli DNAPolymeraseIOligonucleotideextension..........................T7DNAPolymerase;KlenowfragmentPhosphorylation.................................T4PolynucleotideKinase,OptiKinase*Poly(A)tailingofRNA ............................Poly(A)PolymerasePolymerasechainreaction(PCR).....................TaqDNAPolymerase,RubyTaqDNAPolymerase*,FideliTaqDNAPolymerase*,HotStart-ITTaqDNA

Polymerase*Primedsynthesis.................................Klenowfragment,Exonuclease-freeKlenow*,Sequenase*QuantitativeReal-TimePCR ........................VeriQuestqPCRMasterMixes*QuantitativeReal-TimeRT-PCR......................VeriQuestOne-StepqRT-PCRMasterMixes*Replacementsynthesis............................T4andT7DNAPolymeraseRestrictionmapping..............................RestrictionendonucleasesTailingDNA ....................................TerminalTransferaseTailingRNA.....................................Poly(A)PolymeraseTranscription,promoter-specific......................E. coli, SP6,T3andT7RNAPolymerase,GeneScribeIn VitroTranscriptionKit*

Updated(*)fromTabor,S.andStruhl,K.1990“EnzymaticManipulationofDNAandRNA,”Current Protocols in Molecular Biology (F.M.Ausubel,R.Brent,R.E.Kingston,D.D.Moore,J.G.Seidman,J.A.SmithandK.Struhl,eds.),pp.3.5.15.GreenePublishingandWileyInterscience,NewYork.

Applications of Nucleic Acid Modifying Enzymes


Molecular Biology Enzymes | Binding Proteins

70028Y 200 µg70028Z 1,000 µgCustoms by request

Source:E. coli straincontaininganoverproducingcloneofE. coliRecAProtein.

Description:E. coliRecAProtein,encodedbytherecAgeneofE. coli,participatesingeneralrecombination,repairofDNAandregulationofrepair(1,2).Ithasbeenobservedin vitrotohaveDNA-dependentATPaseactivity,promoteproteolyticcleavageofrepressors(3)andtocatalyzeDNAstrandpairingandexchange(4,5).InthepresenceofATP,RecAProteinpromotesthestrandexchangeofsingle-strandDNAfragmentswithhomologousduplexDNA(6).Thisproteinwillformhelicalfilamentswithsingle-strandedandduplexDNA(7,8).

Applications:�� AnalysisofcomplexDNAstructuresbyelectronmicroscopy(9)

�� Site-directedmutagenesis(10)

�� ScreeningofDNAlibraries(11,12)

Properties:MolecularWeight:37.8kDa(13)

Purity:Greaterthan98%pureasdeterminedbySDS-PAGE.Testedforcontaminatingnon-specificendonucle-ases,exonucleases,andribonucleases.

Storage buffer:20mMTris-HCl(pH7.5),1mMDTT,0.1mMEDTA,50%glycerol.

Concentration:5mg/ml,measuredbyA280

(3)

Shipping and storage:Shippedondryice.Storeat-20°C.Forlongtermstoragestoreat-80°C.Avoidrepeatedfreeze/thawcycles.

References:1. Radding,C.M.(1978)Ann.Rev.Biochem.47,

847-880.2. Radding,C.M.(1982) Ann.Rev.Genet.16,405-

437.3. Craig,N.L.andRoberts,J.W.(1981)J. Biol.

Chem. 256,8039-8044.

4. Cox,M.M.andLehman,I.R.(1981)Proc. Natl. Acad. Sci. USA 78,3433-3437.

5. Soltis,D.A.andLehman,I.R.(1983)J.Biol.Chem.258,6073-6077.

6. Radding,C.M.(1991)J.Biol.Chem.266,5355-5358.

7. DiCapua,E.,Engel,A.,Stasiak,A.andKoller,T.(1982)J.Mol.Biol.157,87-103.

8. Register,J.C.andGriffith,J.(1986)Proc.Natl.Acad.Sci.USA83,624-628.

9. Wasserman,S.A.andCozzarelli,N.R.(1985)Proc.Natl.Acad.Sci.USA82,1079-1083.

10.Shortle,D.,Koshland,D.,Weinstock,G.M.andBotstein,D.(1980)Proc.Natl.Acad.Sci.USA77,5375-5379.

11.Honigberg,S.M.,Rao,B.J.andRadding,C.M.(1986)Proc.Natl.Acad.Sci.USA83,9586-9590.

12.Rigas,B.,Welcher,A.A.,Ward,D.C.andWeissman,S.M.(1986)Proc.Natl.Acad.Sci.USA 83,9591-9595.

13.Sancar,A.,StachelekC.,Konigsberg,W.andRupp,W.D.(1980)Proc.Natl.Acad.Sci.USA77,2611-2615.

RecA Protein

70032Y 100 µg70032Z 500 µgCustoms by request

Source:E. coli straincontaininganoverproducingcloneofE. coli SSBprotein.

Description:Single-StrandedDNABindingProteinbindswithhighaffinityinacooperativemannertosingle-strandedDNAanddoesnotbindwelltodouble-strandedDNA(1,2).ItisinvolvedinDNAreplicationandrecombinationin vivo.Thisproteinhasbeenusedtovisualizesingle-strandedDNAbyelectronmicroscopy(3,4,5).Ithasalsobeenusedinconjunc-tionwithRecAproteinforcarryingoutsite-directedmutagenesis(6)andtoselectsequencesfromlibrariesofdouble-strandedDNA(7).SSBmayalsostimulatespecificDNApolymerasesusedinDNAsequencingreactions(8)andhasbeenusedtotargetrestrictionendonucleasedigestionstospecificsitesinsingle-strandedDNAforsubsequentmutagenesis(9).

SSBhasbeenshowntobeeffectiveinfluorescencepolarizationassays(15)andeliminatespausingwhensequencingthroughstrongsecondarystructure.Morerecently,SSBwasusedtohelpobtainlongerreadlengthsinpyrosequencingforSNPanalysis(16,17).

Applications:�� EnhancementofDNApolymeraseactivity�� Fluorescencepolarizationassays(15)

�� AllowsforlongerreadlengthsinpyrosequencingforSNPanalysis(16,17)

�� Eliminatespausingwhensequencingthroughregionsofsingleanddouble-strandedDNAwithstrongsecondarystructure�� Improvesrestrictionenzymedigestions�� Site-directedmutagenesisinconjunctionwithRecA

Properties:Consistsoffouridentical18.9kDasubunits(10,11)

IsoelectricPoint:6.0(18)

NucleotidesBoundPerMonomer:8-16(18)

Thermostability:SSBretainsactivityafter20minutesincubationat65°C.


Storage buffer:50mMTris-HCl(pH7.5),200mMNaCl,0.1mMEDTA,1.0mMDTT,50%glycerol.

Concentration:5µg/µl,measuredbyA280

(12)

Activity:Approximately5µgofSSBproteinisrequiredtopreventopticaldensitychangeof1µgofsingle-strandedDNAuponadditionof10mMMgCl2.

Shipping and storage:Shippedondryice.Storeat-20°C.Forlongtermstorage,storeat-80°C.Avoidrepeatedfreeze/thawcycles.

References:

1. KraussG.,Sindermann,H.,Schomburg,U.andMaass,G.(1981)Biochemistry 20,5346-5352.

2. Weiner,J.H.,Bertsch,L.L.andKornberg,A.(1975)J. Biol. Chem.250,1972-1980.

3. Chrysogelos,S.andGriffith,J.(1982)Proc. Natl. Acad. Sci. USA 79,5803-5807.

4. Sigal,N.,Delius,H.,Kornberg,T.,Gefter,M.L.andAlberts,B.(1972)Proc. Natl. Acad. Sci. USA 69,3537-3541.

5. Muskavitch,K.M.andLinn,S.(1982)J. Biol. Chem. 257,2641-2648.

6. Shortle,D.(1980)Proc. Natl. Acad. Sci. USA 77,5375-5379.

7. Honigberg,S.M.,Rao,B.J.andRadding,C.M.(1986)Proc. Natl. Acad. Sci. USA 83,9586-9590.

8. Kowalczykowski,S.C.,Bear,D.G.andVonHippel,P.H.(1981)inThe Enzymes,3rdedition,ed.P.D.Boyer,(AcademicPress,NewYork)14,373-444.

Single-Stranded DNA Binding Protein (SSB)



Molecular Biology Enzymes | Binding Proteins

Standard concentration, 5 µg/µl70029Y 100 µg70029Z 500 µgHigh concentration, ≥ 10 µg/µl74029Y 300 µg74029Z 1,000 µgCustoms by request

Source:E. coli straincontaininganoverproducingcloneofT4Gene32Protein.

Description:T4Gene32Proteinisasingle-strandedDNAbind-ingproteinwhichisrequiredforT4DNAreplication,recombination,andrepair.Itbindscooperativelytosingle-strandedDNAandin vivo isrequiredinstoichiometricratherthancatalyticquantities(1,2).Italsobindstosingle-strandedRNA(with101-104loweraffinitythansingle-strandedDNA)allowingittocontrolitsownrateofsynthesisattheleveloftranslation(3,4).Ithasbeennotedtoeliminatepausingwhensequencingthroughstrongsecondarystructure.

T4Gene32ProteinhasbeenwidelyusedinstudiesofDNA-proteininteractionsandformarkingregionsofsingle-strandedDNAincytologicalpreparationsviewedbyelectronmicroscopy(5,6).Onaprimedsingle-strandedDNAtemplatetheadditionofT4Gene32Proteinresultsina5to10-foldincreaseintherateofsynthesisbyT4DNAPolymerase(7).T4Gene32ProteinhasbeenshowntobeeffectiveinstimulatingPCR.Inaddition,theenzymehasbeenfoundtoenhanceyieldandprocessivityinRNAamplification(8).

Applications:�� EnhancementofyieldandspecificityinRNAamplification(8)

�� StimulationofPCR(9)

�� Improvingrestrictionenzymedigestions(10)

�� Sequencingthroughregionsofdouble-andsingle-strandedDNApossessingstrongsecondarystructure�� EnhancementofDNApolymeraseactivity(11)

�� IncreaseyieldinRT-PCR(12,13)

Properties:IsoelectricPoint:5.5(14)MolecularWeight:33.5kDaNativeForm:MonomerNucleotidesBoundPerMonomer:10(14)


Storage buffer:20mMTris-HCI(pH8.0),100mMNaCI,1.0mMEDTA,0.5mMDTT,50%glycerol.

Concentrations:PN70029:5µg/µl,measuredbyA280

(15)

PN74029:≥ 10µg/µl,measuredbyA280(15)

Functional Assay: FunctionallytestedinbindingassayofT4Gene32Proteintosingle-strandedDNAasdeterminedbyagarosegelelectrophoresisingel-shiftassay.

Shipping and storage:Shippedondryice.Storeat-20°C.

References:1. Chase,J.W.andWilliams,K.R.(1986)Ann. Rev.

Biochem.55,103-136.2. Sinha,N.K.andSnustad,D.P.(1971)J. Mol. Biol.

62,267-271.3. Krisch,H.M.,Bolle,A.andEpstein,R.H.(1974)

J. Mol. Biol.88,89-104.4. Gold,L.,O’Farrel,P.Z.andRussel,M.(1976)

J. Biol. Chem.251,7251-7262.5. Delius,H.,Mantell,N.J.andAlberts,B.(1972)

J. Mol. Biol.67,341-350.6. Brack,C.,Bickle,T.A.andYuan,R.(1975)J. Mol.

Biol.96,693-702.7. Huberman,J.A.,Kornberg,A.andAlberts,B.M.

(1971)J. Mol. Biol.62,39-52.8. Baugh,L.R.,Hill,A.A.,Brown,E.L.andHunter,

C.P.(2001)Nucl. Acids Res.29 (5),E29.9. Schwarz,K.,Hansen-Hagge,T.andBartram,C.

(1990)Nucleic. Acids Res.18,1079.10.Dombroski,D.F.andMorganA.R.(1985)J. Biol.

Chem.260,415-417.11.Topal,M.D.andSinha,N.K.(1983)J. Biol.

Chem. 258,12274-12279.12.Villalva,C.,Touriol,C.,Seurat,P.,Trempat,P.,

Delsol,G.andBrousset,P.(2001)BioTechniques31,81-86.

13.Jeffries,D.andFarquharson,C.(2002)Anal. Biochem.308,192-194.

14.Kornberg,A.andBaker,T.(1991) DNA Replication, 2ndEdition.

15.Williams,K.R.,Sillerud,L.O.,Schafer,D.E.andKonigsberg,W.H.(1979)J. Biol. Chem.254,6426-6432.

16.Shamoo,Y.,Adari,H.,Konigsberg,W.H.,Williams,K.R.andChase,J.W.(1986)Proc. Natl. Acad. Sci. USA 83,8844-8848.

T4 Gene 32 Protein

9. Milavetz,B.(1989)Nucl. Acids Res.17,3322.10.Sancar,A.,Williams,K.R.,Chase,J.W.andRupp,

W.D.(1981)Proc. Natl. Acad. Sci. USA78,4274-4278.

11.Chase,J.W.andWilliams,K.R.(1986)Ann. Rev. Biochem. 55,103-136.

12.Williams,K.R.,Spicer,E.K.,Lopresti,M.B.,Guggenheimer,R.A.andChase,J.W.(1983)J. Biol. Chem. 258,3346-3355.

13.Griffith,J.D.,Harris,L.D.andRegister,J.(1984)Cold Spring Harbor Symposia on Quant. Biol. 49,553-559.

14.Julin,D.A.,Riddles,P.W.andLehman,I.R.(1986)J. Biol. Chem. 261,1025-1030.

15.Hsu,T.M.,Chen,X., Duan,S.,Miller,R.D.and Kwok,P.-Y.(2001)BioTechniques 31,560-570.

16.Andréasson,A.,Asp,A.,Alderborn,A.,Gyllensten,U.andAllen,M.(2002)BioTechniques32 (1),124-133.

17.Ronaghi,M.(2000)Anal. Biochem286,282-288.

18.Kornberg,A.andBaker,T.(1991)DNA Replication,2ndEdition,328.

Single-Stranded DNA Binding Protein (SSB) (continued from previous page)


Molecular Biology Enzymes | Glycosylases

78310 100 unitsCustoms by request

Source: RecombinantfromGadus morhua(codliver)

Description: Uracil-DNAGlycosylase(UDGorUNG)derivedfromGadus morhua hasalltheattributesoftheenzymederivedfromE. coliwiththeaddedbenefitofbeingheat-labile.Itiscompletelyandirreversiblyinacti-vatedaftera10minuteincubationat50°C.

UDGexcisesuracilfromdU-containingDNAbycleavingtheN-glycosidicbondbetweentheuracilbaseandthesugarbackbone.Thiscleavagegener-atessitesthatareblockedfromreplicationbyDNApolymeraseorpreventedfrombecomingahybridiza-tionsite.

Double-andsingle-strandeddU-containingDNAaresubstratesforUDGwhereasRNAandnormaldT-containingDNAarenot(1).

Uracil-DNAGlycosylase(UDGorUNG)canbeutilizedforthepreventionoffalsepositiveresultsinPCRcausedbytraceamountsofcarry-overcontaminantsfromthepreviousreactions(5).ThiscanbeaccomplishedbysubstitutingdUTPfordTTPinthePCRreactionmixtoincorporatedUTPinallofthePCRproducts.

Pre-incubationofthesubsequentPCRstartingreac-tionswithUDGresultsinexcisionofuracilfromthedU-containingcarry-overcontaminantstopreventamplification.ThetemplatewhichdoesnotcontaindUisunaffectedandwillbeamplifiednormally.UDGisinactivatedduringsubsequentthermocyclingandwillnotaffectthenewdU-richPCRproducts.

Applications:�� StudyofDNArepairandmutationdetection(4)

�� IncreasecloningefficiencyofPCRproducts(2)

�� Increasetheefficiencyofsite-directedmutagenesis(3)

�� Studyofprotein-DNAinteractions(5)

�� Preventionofcarry-overcontaminantsinPCR(6)

Purity:TheenzymeischromatographicallypurifiedandhomogeneousbySDS-PAGE.Testedforcontaminat-ingendonucleasesandexonucleases.

Storage buffer:50mMTris-HCl(pH7.5),100mMNaCl,0.5mMEDTA,1mMDTT,0.1%TritonX-100,50%glycerol.

Assay conditions: Thereactionmixturecontains70mMTris-HCl,pH8.0,10mMNaCl,1mMEDTA,100µg/mlBSA,3H-dUTPlabeledDNA,andUracil-DNAGlycosylase.Incubationisat37°Cfor1hour.

Unit definition:Oneunitistheamountofenzymerequiredtoliber-ate1nmoluracilfromdU-containingDNAinonehourat37°C.

Concentration:1unit/µl


References:1.Lindahl,T.,Ljungquist,S.,Siegert,W.,Nyberg,B.

andSperens,B.(1977)J. Biol. Chem.252,3286-3294.

2.Nisson,P.E.,Rashtchian,A.andWatkins,P.C.(1991)PCR Methods Appl.1,120-123.

3.Kunkel,T.(1985)Proc. Natl. Acad. Sci. USA82,488-492.

4.Vaughan,P.andMcCarthy,T.V.(1998)Nucl. Acids Res.26,810-815.

5.Deuchand,P.R.,McGhee,J.D.andVanDeSande,J.H.(1993)Nucl. Acids Res.21,3437-3443.

6.Longo,M.C.,Berninger,M.S.andHartley,J.L.(1990)Gene93,125-128.

Uracil-DNA Glycosylase, Heat-Labile

71960 400 units2,000 units

Customs by request

Source: E. coli straincontaininganoverproducingcloneofE. coli Uracil-DNAGlycosylase.

Description: Uracil-DNAGlycosylase(UDGorUNG)excisesuracilfromdU-containingDNAbycleavingtheN-glycosidicbondbetweentheuracilbaseandthesugarbackbone.ThiscleavagegeneratesalkalisensitiveapyrimidinicsitesthatareblockedfromreplicationbyDNApolymeraseorpreventedfrombecomingahybridizationsite.

Double-andsingle-strandeddU-containingDNAaresubstratesforUracil-DNAGlycosylasewhereasRNAandnormaldT-containingDNAarenot(1).

Applications:�� StudyofDNArepairandmutationdetection(4)

�� IncreasecloningefficiencyofPCRproducts(2)

�� Increasetheefficiencyofsite-directedmutagenesis(3)

�� Studyofprotein-DNAinteractions(5)

�� Preventionofcarry-overcontaminantsinPCR(6)

Properties:MolecularWeight:31kDaOptimumpH:8.0OptimumTemperature:37°CInactivation:95°Cfor10minutes.Note:Enzymeactivityispartiallyrestoredattem-peratureslowerthan55°C.

Purity:Testedforcontaminatingsingle-anddouble-strandedexonucleasesandendonucleases.

Storage buffer:10mMTris-HCl,pH7.5,50mMKCl,1mMDTT,0.1mMEDTA,0.05%Tween20,and50%glycerol.

Assay conditions:70mMHEPES(pH8.0),50mMEDTA,1mMDTT,0.4µglambdaDNAat37°Cfor30minutes.

Unit definition:Oneunitistheamountofenzymethatcatalyzesthereleaseof60pmolofuracilperminutefromdouble-stranded,uracil-containingDNA.

Concentration:2units/µl

Uracil-DNA Glycosylase Reaction Buffer (1 ml included):200mMTris-HCl(pH8),10mMDTT,10mMEDTA.


References:1.Lindahl,T.,Ljungquist,S.,Siegert,W.,Nyberg,B.

andSperens,B.(1977)J. Biol. Chem.252,3286-3294.

2.Nisson,P.E.,Rashtchian,A.andWatkins,P.C.(1991)PCR Methods Appl.1,120-123.

3.Kunkel,T.(1985)Proc. Natl. Acad. Sci. USA82,488-492.

4.Vaughan,P.andMcCarthy,T.V.(1998)Nucl. Acids Res.26,810-815.

5.Deuchand,P.R.,McGhee,J.D.andVanDeSande,J.H.(1993)Nucl. Acids Res.21,3437-3443.

6.Longo,M.C.,Berninger,M.S.andHartley,J.L.(1990)Gene93,125-128.

Uracil-DNA Glycosylase, E. coli


78334X 500 units78334Y 1,000 units78334Z 2,500 units

Source:E. colistraincontainingamodifiedcloneofT4PolynucleotideKinasethathasfullkinaseactivitybutlacksphosphataseactivity.

Description:OptiKinaseisamodifiedversionofT4PolynucleotideKinase(T4PNK)(1,2,3)whichprovidesgreater32Por33PlabelingefficiencyofDNA.T4PNKisahomotetra-meric,bifunctionalenzymecontaininganN-terminalkinasedomainwhichcatalyzesthetransferoftheg-phosphateofATPtothe5'hydroxylendofpoly-nucleotidesandaC-terminalphosphatasedomainwhichremovesthe3'phosphate.Inmolecularbiologyapplications,T4PNKistraditionallyusedforradiolabellingthe5'-endsofDNAandRNA.How-ever,whenphosphorylatingDNA,T4PNKdisplaysaseverebasebias,asthelabelingefficiencyisvariabledependingonthenucleotideatthe5'-end.Deoxy-guanosineatthe5'-endiskinasedwiththehighestefficiencyfollowedbydA,dT,andthendC,whichiskinasedwiththeleastefficiency(4).Unlikewild-typeT4PNK,OptiKinaseexhibitsnosuchbasebiasordis-crimination(seeFigs.1and2).Also,sinceOptiKinaselacks3'phosphataseactivity,itisparticularlyusefulforlabeling3'phosphorylatedmononucleotidesorRNAwitha3'phosphate.UseOptiKinaseforhigherlevelsofphosphorylationandforgreaterconsistencycomparedtostandardT4PNK.

Applications: �� 5'phosphorylationofsingle-strandedoligonucleotides,bothDNAandRNA�� 5'phosphorylationofdouble-strandedDNA�� EfficientlabelingofDNAwithoutabiastowardthebaseatthe5'end�� 5'phosphorylationofoligonucleotideswitha3'phosphategroup�� 5'phosphorylationofmononucleotideswitha3'phosphategroupthatcangenerateasubstance(pNp)thatcanbeligatedtothe3'endofDNAorRNAbyT4RNALigase

Properties: MolecularWeight:Homotetramerof140kDa

(4x35kDa)OptimumpH:7.6(Tris-HCI)OptimumTemperature:37°C

Purity: Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingexonucleases,endonucle-ases,andribonucleases.

Storage buffer: 20mMTris-HCl(pH7.5),50mMNaCl,1.0mMDTT,0.1mMEDTA,50%glycerol.

Assay conditions: Thereactionmixture(100µl)contains50mMTris-HCI(pH7.6),100µMradiolabeledATP(~1.5x105cpm/nmol),10mMMgCl2,10mM2-mercaptoetha-nol,20µMspermidine,andsubstrate.Incubationisat37°C.

Unit definition: Oneunitistheamountofenzymerequiredtoincor-porate1nmolofphosphatefromradiolabeledATPintoDNAsubstratein30minutesat37°C.

Concentration: 10units/µl

Functional test: >60%phosphorylationofoligonucleotidesubstrate.

Functionally tested 10X OptiKinase Reaction Buffer (1 ml included):0.5MTris-HCI(pH7.5),100mMMgCl2,50mMDTT.

Shipping and storage: Shippedondryice.Storeat-20°C.

References: 1.Wang,L.K.andShumanS.(2001)J. Biol. Chem.

276,26868-26874.2.Wang,L.K.andShumanS.(2002)Nucl. Acids Res.

30,1073-1080.3.Richardson,C.C.(1981)TheEnzymes,3rdedition,

ed.P.D.Boyer,(AcademicPress,NewYork)14,299-314.

4.VanHouten,V.,Denkers,F.,VanDijk,M.,VanDenBredel,M.andBrakenhoff,R.(1998)Anal. Biochemistry265,386-389.

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Molecular Biology Enzymes | Kinases

OptiKinase(PhosphataseminusmutantofT4PNK)

Fig. 2. 5'-Phosphorylation of 5'-C oligo with varying amount of radiolabel. Test5'-Coligonucleotide18-mer(5pmol)andvary-ingamountof33P-gATPwereincubatedwith10unitsofT4PNKorOptiKinasein25μlat37°Cfor30minutes.Thespecificactivitiesoftheresulting5'-phosphorylatedoligonucleotidesweredetermined.

OptiKinase Shifts the Reaction Equilibrium Toward Phosphorylation

5'-Phosphorylation of 5'-C Oligo with Varying Amount of Radiolabel

Fig. 1. 5'-Phosphorylation of single-stranded oligonucle-otides. Testoligonucleotides*(5pmol),identicalexceptattheir5'ends,andanequimolaramountof33P-gATP,weretreatedwith10unitsofT4PNKorOptiKinasein25μlat37°Cfor30minutes.Thespecificactivitiesoftheresulting5'-phosphorylatedoligonucleotidesweredetermined.*Oliognucleotidescorrespondedto31-mers(G,A,T,andC)or18-mers(GGGandCCC).

OptiKinase Overcomes Base Bias5'-Phosphorylation of Single-Stranded Oligonucleotides


Molecular Biology Enzymes | Kinases

70031Y 500 units70031Z 1,000 units70031X 2,500 unitsCustoms by request

Source:E. colistraincontainingacloneofT4PolynucleotideKinase(1).

Description:T4PolynucleotideKinase(PNK)catalyzesthetransferoftheterminalphosphateofATPto5'hydroxylterminiofpolynucleotidessuchasDNAandRNA,oligonucleotides,and3'mononucleotides(2).InthepresenceofADP,itwillalsocatalyzetheexchangeof5'terminalphosphategroupsandATP.Italsopossessesa3'phosphatase(4,5,8,9)and2',3'cyclicphosphodiesteraseactivity(3).Thisenzymeiscom-monlyusedtolabelDNAorRNAwith32Por33Pat5'ends(6).

Applications:�� 5'-labelingofDNAandRNA�� MappingterminiofRNAtranscripts�� Phosphorylationofoligonucleotidesat5'terminipriortoligation�� Removalof3'-phosphorylgroups

Properties:MolecularWeight:140kDa(4x33kDa)OptimumpH:7.6(Tris-HCI)OptimumTemperature:37°COptimumMg2+Concentration:10mMActivators:MgCl2,spermidine,dithiothreitol,

2-mercaptoethanolInhibitors:Inorganicphosphate,pyrophosphate,

ammoniumsulfateInactivation:65°Cfor10minutes

Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingexonucleases,endonucle-ases,andribonucleases.

Storage buffer:50mMTris-HCl(pH7.5),1.0mMDTT,0.1mMEDTA,50%glycerol.

Assay conditions:Thereactionmixture(100µl)contains50mMTris-HCI(pH7.6),100µMradiolabeledATP,10mMMgCl2,10mM2-mercaptoethanol,20µMspermi-dine,andDNAsubstrate.Incubationisat37°C.

Unit definition:Oneunitistheamountofenzymerequiredtoincorporate1nmolofradiolabeledATPintoDNAsubstratein30minutesat37°C.


Functional test:Phosphorylationof10pmolofa17-meroligo-nucleotidewith20pmolradiolabeledATP,>50%incorporationoflabelwithin30minutes.

Functionally tested 10X T4 PNK Reaction Buf-fer (1 ml included):0.5MTris-HCl(pH7.6),100mMMgCl2,100mM2-mercaptoethanol.

T4 PNK Dilution Buffer (1 ml included):50mMTris-HCl(pH8.0).


References:1.Midgley,C.A.andMurray,N.E.(1985)EMBO J.4,

2695-2703.2.Richardson,C.C.(1981)inThe Enzymes,3rd

edition,ed.P.D.Boyer,(AcademicPress,NewYork)14,299-314.

3.Morse,D.P.andBass,B.L.(1997)Biochemistry36,8429-8434.

4.Cameron,V.andUhlenbeck,O.C.(1977)Biochemistry16,5120-5126.

5.Wang,L.K.andShuman,S.(2002)Nucl. Acids Res.30,1073-1080.

6.Maxam,A.M.andGilbert,W.(1980)Methods In Enzymology65,499-560.

7.VanHouten,V.,Denkers,F.,VanDijk,M.,VanDenBrekel,M.andBrakenhoff,R.(1998)Anal. Biochemistry265,386-389.

8.Galburt,E.,Pelletier,J.,Wilson,G.andStoddard,B.(2002)Structure10,1249-1260.

9.Wang,L.K.,Lima,C.D.andShuman,S.(2002)EMBO J.21,3873-3880.

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Shrimp Alkaline Phosphatase78390 500 units 1,000 units 5,000 units

TEMED76320 100 ml

T4 Polynucleotide Kinase (PNK) (E.C.2.7.1.78)


70020Y 200 units70020Z 1,000 unitsCustoms by request

Source:E. colistraincontaininganoverproducingcloneofE. coliDNALigase.

Description:E. coliDNALigase,inthepresenceofNAD,catalyzestheformationofthephosphodiesterbondinduplexDNAcontainingcohesiveends(juxtaposed5'-phosphoryland3'-hydroxyltermini).Also,inthepresenceofcertainmacromoleculessuchasPEG,E. coliDNALigasecatalyzesbluntendligationofDNA(1).TheenzymeispurifiedbytheprocedureofPanasenko,et al.(2).E. coliDNALigasehasbeenemployedinaprocedureforhighefficiencycloningoffull-lengthcDNA(3).

Application:�� Ligationforcloning(1,3)

Properties:MolecularWeight:75kDaOptimumpH:8.1OptimumDivalentCationConcentration:

Mg2+:3mM;Mn2+:6mM

Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleasesandexonucleases.

Storage buffer:10mMTris-HCl(pH7.5),50mMKCl,0.1mMEDTA,10mMammoniumsulfate,1mMDTT,50%glycerol.

Assay conditions:Thereactionmixture(20µl)contains30mMTris-HCl(pH8.0),4mMMgCl2,1mMDTT,26µMNAD,50µg/mlBSA,andλHindIIIfragments.Incubationisat16°Cfor30minutes.

Unit definition:Oneunitistheamountofenzymerequiredtogive50%ligationofλHindIIIfragmentsin30minutesat16°Cwitha5'-DNAterminiconcentrationof0.003µM(7µg/ml)understandardassayconditions.


Functional test:LigationofλHindIIIfragments.Oneunitgives~50%ligationin30minutesat16°C.Verifiedbyagarosegelelectrophoresis.

Functionally tested 10X E. coli DNA Ligase Reaction Buffer (1 ml included):300mMTris-HCl(pH8.0),40mMMgCl2,10mMDTT,0.5mg/mlBSA,260µMNAD.


References:1.Zimmerman,S.B.andPheiffer,B.H.(1983)Proc.

Natl. Acad. Sci.,USA80,5852-5856.2.Panasenko,S.M.,Alazard,R.J.andLehman,I.R.

(1978)J. Biol. Chem.253,4590-4592.3.Okayama,H.andBerg,P.(1982)Mol. Cell. Biol.2,

161-170.4.Panasenko,S.M.,Cameron,J.R.,Davis,R.W.and

Lehman,I.R.(1977)Science196,188-189.

Molecular Biology Enzymes | Ligases

Standard concentration, 1 unit/µl70005Y 100 units70005X 500 units70005 2,000 unitsHigh concentration, 10 units/µl70042X 500 units70042 2,000 unitsCustoms by request

Source:E. colistraincontaininganoverproducingcloneofT4DNALigase.

Description:T4DNALigasecatalyzestheformationofaphos-phodiesterbondbetweenjuxtaposed5'phosphoryland3'-hydroxylterminiinduplexDNAorRNA.Itrepairssingle-strandednicksinduplexDNA,RNA,orDNA/RNAhybridsandwilljoinbothblunt-endandcohesive-endfragmentsofduplexDNAorRNA(1).

ForconvenienceweoffertheLigate-IT™RapidLigationKit(PN78400/10)for5minutesroomtemperatureligation.

Applications:�� Ligationofcohesive-endorblunt-endtermini�� Ligationofsyntheticlinkersoradaptors

Properties:MolecularWeight:55kDaOptimumpH:7.5-7.8OptimumTemperature:16°CforexchangereactionRequirementsforDivalentCation:Mg2+,Mn2+

OptimumMg2+Concentration:10mMSulfhydrylRequirement:2-ME,DTTStimulators:SpermineorSpermidine(0.5to1mM)Inhibitors:Na(>0.2M),K(>0.2M)Inactivation:Heatingat65°Cfor10minutes

Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleases,exonucle-ases,andribonucleases.

Storage buffer:25mMTris-HCl(pH7.6),100mMNaCl,1mMDTT,0.5mMEDTA,50%glycerol.

Assay conditions:T4DNALigaseisassayedaccordingtothepyro-phosphateexchangemethodofWeiss,et al.Thereactionmixture(300µl)contains67mMTris-HCl,(pH7.8),6.7mMMgCl2,10mMDTT,66µMATP,and3.3µMradiolabeledpyrophosphate.Incubationisat37°Cfor20minutes.

Note:ATPisanessentialcofactorforthereaction.ThiscontrastswithE. coliDNALigasewhichrequiresNAD.

Unit definition:OneWeissunitistheamountofenzymerequiredtoconvert1nmolofradiolabeledphosphatefrompyro-phosphateintoabsorbablematerialin20minutesat37°Cunderstandardassayconditions.OneWeissunitequalsabout67cohesiveendligationunits.

Concentration:PN70005:1unit/µl;PN70042:10units/µl

Functional test:ReligationoflinearizedplasmidDNA>80%ofcontrol,asdeterminedbycountingtransformedbacterialcolonies.

Functionally tested 10X T4 DNA Ligase Reac-tion Buffer (1 ml included):660mMTris-HCl(pH7.6),66mMMgCl2,100mMDTT,660µMATP.

T4 DNA Ligase Dilution Buffer (1 ml in-cluded):20mMTris-HCl(pH7.6),60mMKCl,5mMDTT,1mMEDTA,50%glycerol.


References:1.Rossi,R.,Montecucco,A.,Ciarrochi,G.and

Biamonti,G.(1997)Nucl. Acids Res.25(11),2106-2113.

2.Weiss,B.,Jacquemin-Sablon,A.,Live,T.R.,Fareed,G.C.andRichardson,C.C.(1968)J. Biol. Chem.243,4543-4555.

T4 DNA Ligase (ATP-Dependent,E.C.6.5.1.1.)

E. coli DNA Ligase (NAD-Dependent,E.C.6.5.1.2.)


Molecular Biology Enzymes | Ligases

21245 600 units2,500 units

Customs by request

Source:E. colistraincontaininganoverproducingcloneofT4RNALigase.

Description:T4RNALigasecatalyzesligationofa5'-phosphoryl-terminatednucleicacid(donor)anda3'-hydroxylterminatednucleicacid(acceptor)throughtheformationofa3'→5'phosphodiesterbond,withthehydrolysisofATPtoAMPandPPi(1).ItssmallestsubstratesarepNpandNpNpNanditsreactionrate(ligationefficiency)dependsonthekindsofbasesinvolved(1).InadditiontoligatingRNAtoRNA(2),itcanalsoligateDNAtoRNA,althoughslowly(3).TherateofDNAtoDNAligationisextremelyslowandthisligasealmostfailstorecognizedouble-strandednucleicacids.Anoligonucleotidewithbotha5'-phosphateand3'-OHcanserveasbothanacceptorandadonor,yieldingeithercircularormultimericproducts.

Applications:�� LigationofRNAtoRNA�� Circularizationofoligonucleotides(3)

�� Labelingthe3'terminiofRNA(4)

�� Ligationofsingle-strandedDNAtoRNA(5)

�� SpecificmodificationsoftRNAs(6)

�� CircularizationofRNA(7)

�� Cloningoffull-lengthDNA(8)

Purity:Greaterthan90%pureasdeterminedbySDS-PAGE.Testedforcontaminatingdeoxyribonucleasesandribonucleases.

Storage buffer:20mMTris-HCl(pH7.4),50mMNaCl,1.0mMDTT,1.0mMEDTA,50%glycerol.

Unit definition:Oneunitistheamountofenzymethatconverts1nmolofradiolabeledpCpintoitsacid-insolubleformin30minutesat37°C,witholigo(A)nasthesubstrate.


Functional test:Ligationoftwooligonucleotides.

Functionally tested 10X T4 RNA Ligase Reac-tion Buffer (1 ml included):500mMTris-HCl(pH7.5),100mMMgCl2,100mMDTT,10mMATP,0.6mg/mlacetylatedBSA.


References:1.Gumport,R.I.andUhlenbeck,O.C.(1981)in

Gene Amplification and Analysis,eds.Chirikjian,J.G.andPapas,T.S.,(Elsevier,Amsterdam)2,313-335.

2.Romaniuk,P.J.andUhlenbeck,O.C.(1983)Methods in Enzymology100,52-59.

3.Brennan,C.A.,Manthey,A.E.andGumport,R.I.(1983)Methods in Enzymology100,38-52.

4.Uhlenbeck,O.C.andGumport,R.I.(1982)inThe Enzymes,3rdedition,ed.P.D.Boyer,(AcademicPress,NewYork)15,31-60.

5.Tessier,T.C.,Brousseau,R.andVernet,T.(1986)Anal. Biochem.158,171-178.

6.Bruce,A.G.andUhlenbeck,O.C.(1982)Biochemistry21,855-861.

7.White,T.C.andBorst,P.(1987)Nucl. Acids Res.15,3275-3290.

8.Dumas,J.P.,Edwards,J.B.,Delort,J.andMallet,J.(1991)Nucl. Acids Res.19,5227-5233.

T4 RNA Ligase

70440 200 units1,000 units

Source: Recombinant

Description:T4RNALigase2,alsoknownasT4Rnl-2(gp24.1),hasbothintermolecularandintramolecularRNAstrandjoiningactivity.T4RNALigase2ismuchmoreactivethanT4RNALigase1(PN21245)injoiningnicksondouble-strandedRNAthanonjoiningtheendsofsingle-strandedRNA.Theenzymerequiresanadjacent5’phosphateand3’OHforligation.T4RNALigase2canalsoligatethe3’OHofRNAtothe5’phosphateofDNAinadouble-strandedstructure.

Applications:�� IdealforligatingnicksindsRNA�� Ligatingthe3’OHofRNAtothe5’phosphateofDNAinadouble-strandedstructure�� Suitableforligating3’OHorRNAto5’phosphateofDNAinaDNA/RNAhybrid

Purity: Greaterthan90%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleases,exonucle-ases,andribonucleases.

Storage buffer: 10mMTris-HCl,pH7.5,50mMKCl,35mM(NH4)2SO4,0.1mMDTT,0.1mMEDTA,50%glycerol

Assay conditions: Thereactionmixturecontains0.4μgofanequi-molarmixofa23-merand17-merRNAs,T4RNALigase2in1XT4RNALigase2ReactionBufferat37°Cin30minutes(20μlreactionvolume).

Unit definition: Oneunitisdefinedastheamountofenzymerequiredtoligate0.4μgofanequimolarmixofa23-merand17-merRNAat37°Cin30minutes.

Concentration:10units/μl

Functionally tested 10X T4 RNA Ligase 2 Reaction Buffer (1 ml included): 500mMTris-HCl,pH7.5,20mMMgCl2,10mMDTT,and4mMATP.

Storage:Shippedondryice.Storeat-20°C.

T4 RNA Ligase 2


14365 5 mgCustoms by request

Source:Bovinepancreas

Description:DNaseIisanendonucleasethatcatalyzesthedeg-radationofbothsingleanddouble-strandedDNAtoyielddi-,tri-,andoligonucleotideswith5'-phosphateand3'-hydroxyltermini(1-4).DNaseIisstabilizedbytheadditionofCa2+andrequiresadivalentmetalcationforactivity.Itrandomlyproducesnicksindouble-strandedDNAinthepresenceofMg2+,butinthepresenceofMn2+,bothstrandsofdouble-strandedDNAarecleaved,leavingblunt-endfragmentsorthosewithoneortwoprotrudingnucleotides(1,2).

Applications:�� LabelingofDNAbynicktranslationwithDNAPolymeraseI(PN70010Y)�� RemovalofDNAfromproteinpreps

Properties:MolecularWeight:31kDaOptimumpH:7.0,moststableatpH5.0-6.0.Activators: Mg2+andCa2+formaximumactivation. Mg2+forindependentstrandcleavage,random

sites. Mn2+forsimultaneouscleavageofbothstrandsto

yieldblunt-endedand1-2nucleotideprotrudingfragments.

Inhibitor:EDTAorEGTAHeatInactivation:75°Cfor10minutes.AddEDTA

toexcessrelativetoMg2+beforeheatingtoavoidchemicalscissionofRNA.

Purity:Testedforcontaminatingribonucleases.

Form:Lyophilizedpowderwithglycinestabilizer.

Activity:≥2,000units/mgdrybasis

Unit definition:Oneunitcausesanincreaseinabsorbanceat260nmof0.001perminutepermlat25°C,pH5.0,whenactingonhighlypolymerizedDNAinthepres-enceofionizedmagnesiumandcalcium(3).

Shipping and storage:Shippedondryice.Storeat4°C.

References:1.Bernardi,A.,Gaillard,C.andBernardi,G.(1975)

Eur. J. Biochem.52,451-457.2.Clark,P.andEichhorn,G.L.(1974)Biochemistry

13,5098-5102.3.Kunitz,M.(1950)J. Gen. Physiol.33,349-363.4.Melgar,E.andGoldthwait,D.A.(1968)J. Biol.

Chem.243,4409-4416.

Molecular Biology Enzymes | Nucleases—DNases

DNase I, Lyophilized (E.C.3.1.21.1)

14367 100 units500 units

Customs by request


Description:DNaseIisanendonucleasethatcatalyzesthedegra-dationofbothsingle-anddouble-strandedDNAtoyielddi-,tri-,andoligonucleotideswith5'-phosphateand3'-hydroxyltermini(1-4).DNaseIisstabilizedbytheadditionofCa2+andrequiresadivalentmetalcationforactivity.Itrandomlyproducesnicksindouble-strandedDNAinthepresenceofMg2+,butinthepresenceofMn2+,bothstrandsofdouble-strandedDNAarecleaved,leavingblunt-endfragmentsorthosewithoneortwoprotrudingnucleotides(1,2).

Applications:�� DegradationoftemplateDNAfromRNAafterin vitrotranscription�� DeterminefootprintsofDNAbindingproteins�� NicktranslationwithDNAPolymeraseI(PN70010Y)�� RemovalofDNAfromRNApreparationspriortoRT-PCR(5)

�� GenerationofrandomoverlappingDNAlibraries(6)

Properties:MolecularWeight:31kDaOptimumpH:7.0,moststableatpH5.0-6.0.Activators: Mg2+andCa2+formaximumactivation. Mg2+forindependentstrandcleavage,random

sites. Mn2+forsimultaneouscleavageofbothstrandsto

yieldblunt-endedand1-2nucleotideprotrudingfragments.

Inhibitor:EDTAorEGTAHeatInactivation:75°Cfor10minutes.AddEDTA

toexcessrelativetoMg2+beforeheatingtoavoidchemicalscissionofRNA.

Purity:Testedforcontaminatingribonucleasesandproteases.

Storage buffer:1mMCaCl2and50%glycerol.

Unit definition:Oneunitincreasestheabsorbanceat260nmby0.001perminutepermlat25°CandpH5.0whenactingonhighlypolymerizedDNAinthepresenceofionizedmagnesiumandcalcium(3).

Concentration:Approximately2units/µl

Shipping and storage:Shippedondryice.Recommendedstoragetem-peratureis4°C;howeverthismaterialisalsostablewhenstoredat-20°C.

References:1.Bernardi,A.,Gaillard,C.andBernardi,G.(1975)

Eur. J. Biochem.52,451-457.2.Clark,P.andEichhorn,G.L.(1974)Biochemistry

13,5098-5102.3.Kunitz,M.(1950)J. Gen. Physiol.33,349-363.4.Melgar,E.andGoldthwait,D.A.(1968)J. Biol.

Chem.243,4409-4416.5.Kienzle,N.,Young,D.,Zehntner,S.,Bushell,G.and

Sculley,T.B.(1996)BioTechniques20,612-616.6.Anderson,S.(1981)Nucl. Acids Res.9,3015-

3027.

DNase I, Solution (E.C.3.1.21.1)



78411 1,000 units 2,500 units

Source:Recombinant

Description:RecombinantDNaseIisoverexpressedandpurifiedfromanon-animalhost,Pichia pastoris,whichisvastlylowerinendogenousRNaselevelsthanpan-creatictissue,thesourceforbovineDNaseI.Hence,therecombinantenzymeisreadilypurifiedwithoutanydetectableRNasecontamination(Fig.1).

DNaseIisanendonucleasethathydrolyzesphosphodiesterlinkagesinDNAtoyielddi-,tri-,andoligonucleotideswitha5'-phosphateanda3'-hydroxyltermini(1).DNaseIcancleavedsDNA,ssDNA,chromatin,andDNA:RNAhybrids.However,thecleavageratesforssDNAandDNA:RNAhybridsaremuchlowerthandsDNA.ThisenzymeistotallyinactiveagainstRNA(2).Formaximumactivity,DNaseIrequiresbothCa2+andMg2+(3).Ca2+stabilizesDNaseItomaintaintheactiveconformationandMg2+isrequiredforactivity.

Applications:�� RemovalofDNAfromproteinandRNApreparations�� DegradationoftemplateDNAfromRNAafterin vitrotranscription�� RemovalofgenomicDNApriortoRT-PCR�� NicktranslationwithDNAPolymeraseI(PN70010Y)

Properties:MolecularWeight:~30kDaActivators:Mg2+andCa2+formaximumactivityInhibitors:EDTAandEGTAHeatInactivation:75°Cfor10minutesata

0.1unit/µlDNaseIconcentration.AddEDTAtoafinalconcentrationof5mMbeforeheatingtoavoidchemicalscissionofRNA.

Purity:Testedforcontaminatingribonucleasesandprote-ases.ResidualDNAcontaminationwastestedbyrealtimequantitativePCR.

Storage buffer:20mMTris-HCl,pH7.6,50mMNaCl,2mMCaCl2,2mMMgCl2,1mMDTT,0.1mg/mlpefablocSC,and50%glycerol.

Assay conditions:100µgcalfthymusDNAisincubatedin1Xreactionbufferwith40to70unitsofrDNaseIat25°C.Theabsorbanceincreaseismeasuredat260nm.

Unit definition:Oneunitistheamountofenzymethatcausesanabsorbanceincreaseof0.001perminutein1mlat260nmat25°C.


10X rDNase I Reaction Buffer (1 ml included): 400mMTris-HCl,pH7.9,100mMNaCl,60mMMgCl2,10mMCaCl2.

Stop Solution (1 ml included): 50mMEDTA,pH8.0.


References:1.Kunitz,M.(1950)J. Gen. Physiol.33,349.2.Sutton,D.H.,Conn,G.L.,Brown,T.,and

Lane,A.N.(1997)Biochem. J.321,481-486.3.Clark,R.andEichhorn,G.L.(1974)Biochem.13,

5098.

rDNase I, RNase-Free

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Fig. 2. Effectiveness of rDNase I to remove DNA.1µgofhumanHeLacelltotalRNAand1µgoflambdaDNAwereco-incubatedwithout(-)orwith(+)2unitsrDNaseIin1XrDNaseIReactionBufferina10µlreactionvolume.Following15minutesat22°C,1µlofStopSolutionwasaddedandthereactionswereheat-inactivatedat65°Cfor10minutesbeforeloadingona1%agarosegel.ThisdemonstratesefficientremovalofDNAbut not RNA.

Fig. 1. Absence of RNase activity in rDNase I.2µgofhumanHeLacelltotalRNAwasincubatedwithout(-)orwith(+)2unitsrDNaseIin1XrDNaseIReactionBufferina10µlreactionvolume.Following15minutesat22°C,1µlofStopSolutionwasaddedandthereactionswereheat-inactivatedat65°Cfor10minutesbeforeloadingona1%agarosegel.TheintegrityoftheribosomalRNAindicatesthatnoRNAdigestiontookplace.



78314 100 units500 units

Customs by request

Source:Pichia pastorisstraincontainingoverproducingcloneofPandalus borealisDNase.

Description:ShrimpDNaseisanendonucleasethatcleavesphosphodiesterlinkagesinDNAtoyielddi-andoligonucleotideswith5'-phosphateand3'-hydroxyltermini.ThisDNasehasaremarkablyhighspecificactivitytowardsdouble-strandedDNA(dsDNA).TheactivitytowardsdsDNAis5,000-foldhigherthantowardssingle-strandedDNAandthuscanbeusedselectivelytodegradedsDNA,leavingsingle-strand-edDNAintact.TheactivityofthisenzymedependsonMg2+concentration(Fig.1)andisstimulatedbyCa2+.However,Ca2+alsostimulatestheRNaseactiv-ityofShrimpDNaseandshouldbeavoidedwhenRNAintegrityiscritical.

TheDNaseactivityfavorslowionicstrength.Activ-itydecreaseswithincreasingionicstrength.Thisrecombinantenzymecanbeheat-inactivatedbyamoderateheattreatmentwithouttheuseofEDTA(Fig.2).ShrimpDNaseistotallyinactivatedat70°Caftera25-30minutesincubation.

Applications:�� SelectivedegradationofdsDNAleavingssDNAandRNAintact�� RemovalofDNAfromRNApriortoRT-PCR�� RemovalofDNAtemplateafterin vitrotranscription�� NicktranslationwithDNAPolymeraseI(PN70010Y)�� FootprintdeterminationofDNAbindingprotein�� Usedinremovalofcarry-overcontaminantsinPCR(1)

Properties:MolecularWeight:47kDAOptimumpH:8.0Km(apparent):0.01mg/mlActivators:Mg2+(Optimum:10mM)andCa2+

(Optimum:1mMinthepresenceofMg2+)Inactivation:70°Cfor30minutesIonicStrength:Low(10-20mMTris-HCl)

Purity: Greaterthan98%pureasdeterminedbySDS-PAGE.Testedforcontaminatingribonucleaseandproteases.

Storage buffer: 20mMTris-HCl,pH7.5,2mMMgCl2,10mMNaCland50%glycerol.

Assay conditions:Thereactionmixturecontains100mMNaAC,pH5.0,5mMMgCl2,50µg/mlcalfthymusDNAandShrimpDNase.Incubationtemperatureis25°C(1mlreactionvolume).

Unit definition: Oneunitincreasestheabsorbanceat260nmby0.001O.D.perminuteat25°CandpH5.0usinghighmolecularweightDNAasasubstrateaccordingtothemethodofKunitz(2).



References:1.PCRCarryOverPreventionMethodiscoveredby

USpatent6,541,204andequivalents.AlicenseforusingthepatentedmethodisconveyedbypurchaseofShrimpDNasefromAffymetrix.

2.Kunitz,M.(1950)J. Gen. Physiol.33,349-363.

Shrimp DNase, Recombinant

Fig. 2. Residual activity of Shrimp DNase.60unitsShrimpDNasein200µlassaybufferwasincubatedat70°C.Aliquotsweretakenoutatindicatedintervalsandresidualactivitywasmeasured.ShrimpDNasewastotallyinactivatedat70°Cin25to30minutes.

Fig. 1. DNase activity as a function of [Mg2+] in Tris-buffer at pH 8.0.Minimumof2mMmagnesiumwasrequiredforDNaseactivity.Highestactivitywasobservedat10mM(datanotshown).

78331 1 ml

Description: RecommendedforuseintheProkaryoticTargetPreparationprotocolfromAffymetrixfortheGeneChip®P. aeruginosaGenomeArrayandtheGeneChipE. coliAntisenseGenomeArray.

10X DNase I Buffer Formulation: 100mMTrisAcetate,pH7.5,100mMMagnesiumAcetate,and500mMPotassiumAcetate

Storage:4°C

10X DNase I Buffer

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Molecular Biology Enzymes | Nucleases—Endonuclease

78300 50,000 unitsCustoms by request

Source:E. coli straincontaininganoverproducingcloneoftheT4EndonucleaseVIIprotein.

Description:T4EndonucleaseVIIisaDNAjunctionspecificendo-nuclease(alsoreferredtoasaresolvaseoracleav-ase).T4EndonucleaseVIIistheproductofgene49ofphageT4.Thisgeneproducthas157aminoacidresiduesandamassof18kDa.T4EndonucleaseVIIfunctionsin vivotoresolvebranchedDNAstructuresinnewlysynthesizedDNA.ItdoesthisbycleavingtheDNAflankingthebranchpriortorearrangementandreannealingoftheDNAstrands.T4Endonucle-aseVIIalsorecognizesmismatches,insertionordeletionloops,gaps,andapurinic/apyrimidinicsites,andcreatesnicksintheDNAstrandsatthesesites(1).ThewidevarietyofsubstratesacceptedsuggeststhatT4EndonucleaseVIIrecognizeschangesinthestructureorconformationofDNAratherthanbind-ingtoaspecificsequence(2).T4EndonucleaseVIIhasbeencrystallizedasadimer(3),althoughthenumberofsubunitsrequiredforbranchresolutionisunclear,asmultiplesofdimershavebeenreportedtobindDNAbranchstructuresduringin vitroDNAbindingexperiments(4).

Applications:�� SNPanalysis(5,6)

�� Mutationdetection(5,6)

Storage buffer:20mMTris-HCI(pH8.0),5mMDTT,50%glycerol.


Functional assay: 1500units(15ng)ofT4EndonucleaseVIIdegrades50%of100fmolTAMRAlabeleddouble-stranded24-meroligonucleotidesubstratein15minutesat37°Cin50mMTris-HCl,pH8.0,10mMMgCl2,10mMβ-MEand0.5µg/µlBSA.

T4 Endonuclease VII Dilution Buffer (1 ml included): 10mMTris-HCI(pH7.5),1mMEDTA,50%glycerol.


References:1.Kemper,B.(1997)inDNA Damage and Repair,

eds.Nickoloff,J.A.,Hoekstra,M.(HumanaPress,Totowa,NJ),1,179-204.

2.Greger,B.andKemper,B.(1998)Nucl.Acids Res.26,4432-4438.

3.Raaijmakers,H.,Vix,O.,Toro,I.,Golz,S.,Kemper,B.andSuck,D.(1999)Embo J.,18,1447-1458.

4.Kupfer,C.,Lee,S.andKemper,B.(1998)J.Biol.Chem.273,31637-31639.

5.Youil,R.,Kemper,B.andCotton,R.G.(1995)Proc. Natl. Acad. Sci. USA 92,87-91.

6.Golz,S.andKemper,B.(1999)Nucl.Acids Res.27,e7.

Related products – Binding protein

Single-Stranded DNA Binding Protein70032Y 100 µg 70032Z 500 µg

T4 Gene 32 ProteinStandard concentration, 1-5 µg/µl70029Y 100 µg 70029Z 500 µgHigh concentration, ≥ 10 µg/µl74029Y 300 µg74029Z 1,000 µg

T4 Endonuclease VII (T4gp49,HJResolvase)

78454 1,000 units 5,000 units

Source:AnE. colistrainwhichcarriestheclonedhumanAPE1gene.

Description: Humanapurinic/apyrimidic(AP)endonuclease(APE1)cleavesthephosphodiesterbackboneaDNAstrandimmediately5'toanAPsiteviaahydrolyticmechanism.Cleavagegeneratesasingle-strandDNAbreakwitha3'-hydroxyland5'-deoxyribosephosphateterminus.Inadditiontoendonucleaseactivity,APE1hasbeenreportedtohaveweakDNA3'-diesterase,3'to5'exonuclease,andRNaseHactivities.APE1shareshomologywithEscherichia coliexonucleaseIIIprotein,andisalsoreferredtoasHAP1orRef-1.

Applications:�� Singlecellgelelectrophoresis(cometassay)�� Alkalineelution�� Alkalineunwinding�� Modifiednicktranslation�� Mismatchrepair

Purity: Testedforcontaminatingendonucleases,exonucle-ases,andribonucleases.

Storage buffer:10mMTris-HCl(pH7.5),50mMKCl,1mMDTT,0.1mMEDTA,0.05%Tween-20,50%Glycerol


Unit definition:Oneunitisdefinedastheamountofenzymerequiredtocleave20pmolofanoligonucleotideduplexcontainingasingleAPsiteinatotalreactionvolumeof10µlin1hourat37°C.AnAPsiteiscreatedbytreating20pmolofanoligonucleotideduplexcontainingasingleuracilresiduewith1unitofE.coliUracil-DNAGlycosylase(UDGorUNG,PN71960)for2minutesat37°C.

10X APE 1 Reaction Buffer (1 ml included):200mMTris-acetate(pH8.0),500mMpotassiumacetate,100mMmagnesiumacetate,10mMDTT.

Storage:Shippedondryice.Storeat-20°C.

References:1.Demple,B.et al.(1991)Proc. Natl. Acad. Sci. USA

88,11450-11454.2.Barzilay,G.et al.(1995)Nucl. Acids Res.,23,

1544-1550.3.Barzilay,G.et al.(1995)Nature Struc. Biol.2,

451-468.4.Robson,C.N.andHickson,D.I.(1991)Nucl. Acids

Res.19,5519-5523.5.Flaherty,D.M.(2001)Am. J. Respir. Cell. Mol. Biol.

25,664-667.

Human Apurinic/Apyrimidinic Endonuclease 1 (APE 1)


Standard concentration, 10 units/µl70073Z 2,500 units70073X 5,000 unitsHigh concentration, 20 units/µl72073 5,000 unitsCustoms by request

Source:E. colistraincontaininganoverproducingcloneofE. coliExonucleaseI.

Description:ExonucleaseIhydrolyzessingle-strandedDNAinthe3'→5'direction,releasing5'-mononucleotidesandleavingtheterminal5'-dinucleotideintact.Hydrolysisisprocessiveandcannotproceedifthe3'terminusisphosphorylated.ExonucleaseIcanbeusedtomeasuretheendonucleolyticcleavageofcovalentlyclosedcircularsingle-strandedDNAreactedwithanendonucleaseofinterest(1).Inaddition,DNAhelicaseactivitycanbemeasuredutilizingExonucleaseI(2).

ExonucleaseIisparticularlyusefulinpreparingtheproductsofPCRforapplicationsinvolvingsequencingorlabelingmethods(3).Typically,theexcessprimersandanyotherextraneoussingle-strandedDNApresentinPCRproductswillinterferewithsubsequentenzymaticreactionsinvolvingDNAsynthesis.ThehydrolyticpropertiesofExonucleaseIdegradeallsingle-strandedDNApresentinthePCRmixtureallowingtheproducttobeusedmoreefficientlyinotherapplications.WhencombinedwithShrimpAlkalinePhosphatase(PN78390)fordNTPdephosphorylation,theuseofalternativepurificationmethods,suchascolumns,gelsormagneticseparations,arecompletelyeliminated.

Applications:�� Eliminationofresidualsingle-strandedDNAcontaininga3'terminus�� MeasuringendonucleolyticcleavageofcovalentlyclosedcircularssDNA�� MeasuringDNAhelicaseactivity

Properties:MolecularWeight:55kDaHeatInactivation:80°for15minutesDegradestoterminaldinucleotidesDegradesglycosylatedDNAOptimumTemperature:37°C

Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleases,double-strandedexonucleases,andribonucleases.

Storage buffer:20mMTris-HCI(pH7.5),5mM2-mercaptoethanol,0.5mMEDTA,50%glycerol.

Assay conditions:Thereactionmixture(100µl)contains67mMglycinebuffer(pH9.5),10mM2-mercaptoetha-nol,6.7mMMgCl2,0.5mMdenaturedDNA,andenzyme.Incubationisat37°Cfor30minutes.

Unit definition:Oneunitistheamountofenzymewhichcatalyzesthereleaseof10nmolofacid-solublenucleotidefromdenaturedDNAin30minutesat37°Cunderstandardconditions.

Concentration:PN70073Z/X:10units/µl;PN72073:20units/µl

Functional assay:TreatedPCRproductwithExonucleaseItodegradeunincorporatedprimersbeforesequencingwithSequenaseVersion2.0DNASequencingKit(PN70770).


References:1.Goldmark,P.J.andLinn,S.(1972)J. Biol. Chem.

247,1849-1860.2.Rosamond,J.,Telander,K.M.andLinn,S.(1979)

J. Biol. Chem.254,8646-8652.3.Werle,E.,ScneiderC.,Renner,M.,Völker,M.and

Fiehn,W.(1994)Nucl. Acids Res.22,4354-4355.4.Hanke,M.andWink,M.(1994)BioTechniques

17,858-860.

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Exonuclease I

Molecular Biology Enzymes | Nucleases—Exonucleases

Exonuclease comparison

Exonuclease Gene(s) DNA substrates Mode of action Acid-soluble products

ExonucleaseI sbcB(xonA) Single-stranded 3'→5' 5'-dNMP5'-terminalpNpN

ExonucleaseIII xthA Duplex 3'→5' 5'-dNMP

ExonucleaseVII xseA, xseB Single-stranded3'→5'5'→3' Oligonucleotides

T7Gene6Exonuclease T7 gene 6 Duplex 5'→3' Oligonucleotides5'-dNMP


70023Y 5,000 units70023Z 25,000 unitsCustoms by request

Source:E. colistraincontaininganoverproducingcloneoftheE. coli xthAgene(ExonucleaseIII).

Description:ExonucleaseIII,themajorapurinic/apyrimidinic(AP)endonucleaseinE. coli(1),isa3'-5'exonucleasespecificfordsDNA.Thisenzymecatalyzesthestep-wiseremovalof5'-mononucleotidesfromthe3'-endsofdsDNAbutnotfromssDNA.ExonucleaseIIIisactiveondsDNAwithbluntends,5'-overhangs,andnicks,butnotonprotruding3'-ends4basesorlonger(2).Thesepropertiesareutilizedtofacilitatestrand-specificlabeling,unidirectionaldeletions,andselectivestranddegradation.ExonucleaseIIIalsohasseveralotheractivitiesincluding:digestionoftheRNAstrandofanRNA-DNAheteroduplex(3),strand-cleavageonthe5'sideofAPsitesinbothdsDNAandssDNA(4),removalof3'-phosphatesfromdsDNA(5),andanincreaseinMutYturnover(6).

Applications:�� Preparingstrand-specificradioactiveprobeswithaDNApolymerase�� Preparingsingle-strandedDNAtemplatesfordideoxysequencing(3)

�� Constructingunidirectionaldeletions�� LongPCR�� DNAfootprinting�� DNArepair/mutationdetectionstudies

Properties:MolecularWeight:32kDaInactivation:Heatat75°Cfor10minutesoradd8µl

0.5MEDTAina200µlreactionvolume.OptimumpH:7.6OptimumTemperature:37°COptimumMg2+Concentration:5mM

Purity:Greaterthan99%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleasesandribonuclease.

Storage buffer:50mMTris-HCl(pH8.0),1mMDTT,100mMKCl,50%glycerol.

Assay conditions:Thereactionmixture(200µl)contains50mMTris-HCl(pH8.0),10mM2-mercaptoethanol,5mMMgCl2,activatedDNA,andenzyme.Incubationisat37°Cfor30minutes.

Unit definition:Oneunitwillcatalyzethereleaseof1nmolofnucleotidestoacidsolubleformin30minutesat37°Cunderstandardassayconditions.


Functional test:Productionofaseriesofdeletionsfromalinearizedplasmid,witha4base3'protrusionatoneendanda5'overhangorbluntendattheother.

Functionally tested 10X Exonuclease III Reaction Buffer (1 ml included):660mMTris-HCl(pH8.0),66mMMgCl2,50mMDTT.


References:1.Taylor,A.F.andWeiss,B.(1982)J. Bact.151(1),

351-357.2.Henikoff,S.(1984)Gene28,351-359.3.Linn,S.M.(1982)inLinn,S.M.andRoberts,R.J.

ed.Nucleases,291-309,ColdSpringHarborLaboratoryPress.

4.Shida,T.,Noda,M.andSekijuchi,J.(1996)Nuc. Acids Res.24(22),4572-4576.

5.Doetsch,P.W.andCunningham,R.P.(1990)Mutat Res.236(2-3),173-201.

6.Pope,M.A.,Porello,S.andDavid,S.(2002)J. Biol. Chem.277(25),22605-22615.

Exonuclease III (E.C.3.1.11.2)


Source:E. colistraincontainingoverproducingclonesencod-ingboththelarge(xseA)andsmall(xseB)subunitsofExonucleaseVII.

Description:ExonucleaseVIIisastrictsingle-stranddirecteden-zymewith5'→3'and3'→5'exonucleaseactivities,makingittheonlybi-directionalexonucleasewithsingle-strandedspecificity(1,2).ExonucleaseVIIhasnoapparentrequirementfordivalentcations,asitisfullyactiveinthepresenceofEDTA.Initialreactionproductsareacidinsolubleoligonucleotideswhicharefurtherhydrolyzedintoacidsolubleform.Theproductsoflimiteddigestionsaresmalloligomers(dimerstododecamers).

Applications:�� MappingpositionsofintronsingenomicDNA(3)

�� RemovalofvectorDNAfrominsertswithpoly(dA-T)tails(4)

�� RemovalofexcessPCRprimers(5)

�� Removalofsingle-strandedover-hangsproducedbyrestrictionendonucleases

Properties:MolecularWeight: xseAsubunit=51.8kDa; xseBsubunit=8.8kDaOptimumTemperature:37°COptimumpH:8.0Inactivation:95°Cfor10minutes

Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleases,double-strandedexonucleases,andribonucleases.

Storage buffer:50mMTris-HCl(pH8.0),200mMNaCl,0.5mMEDTA,10mMDTT,50%glycerol.

Assay conditions:Thereaction(50µl)contains50mMTris-HCl(pH7.9),50mMpotassiumphosphate(pH7.6),8.3mMEDTA,10mM2-mercaptoethanol,denaturedDNA,andenzyme.

Unit definition:Oneunitistheamountofenzymerequiredtocatalyzetheconversionof1nmolofnucleotidetoacid-solublenucleotidein30minutesat37°Cunderstandardassayconditions.



References:1.ChaseJ.W.andRichardson,C.C.(1974)J. Biol.

Chem.249,4545-4552.2.ChaseJ.W.andRichardson,C.C.(1974)J. Biol.

Chem.249,4553-4561.3.Berk,A.J.andSharp,P.A.(1978)Proc. Natl. Acad.

Sci. USA75,1274-1278.4.Goff,S.P.andBerg,P.(1978)Proc. Natl. Acad. Sci.

USA75,1763-1767.5.Li,H.H.,Cui,X.andArnheim,N.(1991)Nucl.

Acids Res.19,3139-3141.

Exonuclease VII

Molecular Biology Enzymes | Nucleases—Exonuclease


Molecular Biology Enzymes | Nucleases—Exonucleases


Source:E. coli straincontaininganoverproducingcloneofT7Gene6Exonuclease(1).

Description:T7Gene6ExonucleasehydrolyzesduplexDNAnon-processivelyinthe5'→3'directionfromboth5'-phosphorylor5'-hydroxylnucleotidesbyliberat-ingoligonucleotides(2)aswellasmononucleotidesuntilabout50%oftheDNAisacidsoluble(1,3).Italsodegradesnucleotidesatthegapsandnicksofdouble-strandedDNAfromthe5'-terminiandRNAfromRNA:DNAhybridsinthe5'→3'direction.

T7Gene6ExonucleaseissimilartoLambdaExonu-cleaseinthatitcatalyzesthestepwisehydrolysisofduplexDNAfromthe5'terminiliberating5'mono-nucleotides.However,unlikeLambdaExonuclease,theenzymehaslowprocessivityanditwillremoveboth5'-hydroxyland5'-phosphoryltermini(4).

ItalsodegradesRNAandDNAfromRNA:DNAhy-bridsinthe5'→3'directionbutisunabletodegradeeitherdouble-orsingle-strandedRNA.

Applications:�� Controlledstepwisedigestionofdouble-strandedDNAfromthe5'termini�� GeneratingssDNAtemplatesforsequencing(5)orSNPanalysis(6)

Properties:MolecularWeight:32kDaOptimumpH:7.5OptimumTemperature:37°CInactivation:75°Cfor10minutesoradd2µlof

0.5MEDTAfora50µlreactionvolume.

Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleasesandribonucleases.

Storage buffer:50mMKPO4(pH6.5),1mMEDTA,1mMDTT,50%glycerol.

Assay conditions:Thereactionmixture(50µl)contains50mMTris-HCI(pH8.1),5mMMgCl2,20mMKCI,5mM2-mercap-toethanol,double-strandedDNA,andenzyme.Incubationisat37°Cfor15minutes.

Unit definition:Oneunitistheamountofenzymerequiredtorelease1nmolofacidsolublenucleotidein15minutesat37°Cunderstandardassayconditions.


Functional test:Conversionof0.5pmolofλDNAtosingle-strandedhalfmoleculesby75unitsofenzymein30minutesat37°C.Verificationbyagarosegelelectrophoresis.

Functionally tested 5X T7 Gene 6 Exonuclease Reaction Buffer (1 ml included):200mMTris-HCl(pH7.5),100mMMgCl2,250mMNaCl.


Digestion protocolDigestion of double-stranded DNA with T7 Gene 6 ExonucleaseDNA(~2.5μg/μl) ___μl5XT7Gene6ExonucleaseBuffer 3μlT7Gene6Exonuclease(5units/μgDNA) ___μlWater to15μl

Incubateat37°Cfor30minutes.

Incubateat80°Cfor10minutestoinactivatetheenzyme.

Productsshouldbecheckedonanagarosegel.ToavoidtrappingofDNAinthewellsofagarosegelsor“sticky-end”artifactbands,heatsamplesto65–75°Cfor3–4minutespriortoloading.

Note: Theabovebufferisa“mediumsalt”bufferwhichworksformanyrestrictionenzymesbutmaynotworkforallofthem.Ifaspecificbufferisusedfordigestion,subsequentdigestionwithT7Gene6Exonucleaseusuallystillworkssatisfactorily.Subsequentreactionsshouldbecarriedoutintheappropriatebuffer.Ethanolprecipitationafterdiges-tionmaybeusedtochangebuffers.

References:1.Kerr,C.andSadowski,P.D.(1972)J. Biol. Chem.

247,311-318.2.Kornberg,A.andBaker,T.(1991)DNAReplication,

SecondEdition,591.3.Thomas,K.R.andOlivera,B.M.(1978)J. Biol.

Chem.253,424-429.

4.Ausubel,F.M.,Brent,R.,Kingston,R.E.,Moore,D.D.,Seidman,J.G.,Smith,J.A.andStruhl,K.,(1987)Current Protocols in Molecular Biology (JohnWileyandSons,Inc.).

5.Shon,M.,Germino,J.andBastia,D.(1982)J. Biol. Chem.257,13823-13827.

6.Nikiforov,T.T.,Rendle,R.B.,Goelet,P.,Rogers,Y.H.,Kotewicz,M.L.,Anderson,S.,Trainor,G.L.andKnapp,M.R.(1994)Nucl. Acids Res22,(20),4167-4175.

7.Engler,M.J.andRichardson,C.C.(1983)J. Biol. Chem.258,11197-11205.

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T7 Gene 6 Exonuclease


27-0330-02 1 gmCustoms by request


Description:RNaseIcatalyzesthehydrolysisof3'→5'phos-phodiesterbondsofRNAwiththeformationofoli-goribonucleotidesterminatinginpyrimidine2'→3'cyclicphosphate.RNaseIisapreparationcontainingamixtureof70%RNaseA.Theremaining30%consistsofotherisozymesofRNase.

Applications:�� RemovalofRNAfromDNA�� RemovalofRNAfromproteinpreparations(2)

Form:Lyophilizedpowder

Properties:Inhibitors:Ribonucleaseinhibitorandvanadyl-

ribonucleosidecomplexes.Inactivation:70°Cfor15minutes

Purity:Theenzymeischromatographicallypurifiedandistestedforcontaminatingproteases.

Unit definition:One(Kunitz)unitistheamountofenzymewhich

catalyzesthehydrolysisofyeastRNAtoyieldafirstordervelocityconstantof1.0at25°C,pH5.0(1).

Concentration:Approximately70Kunitzunits/mg


References:1.Asubel,F.M.,Brent,R.,Kingston,R.E.,Moore,

D.D.,Seidman,J.G.,Smith,J.A.andStruhl,K.(1998)Current Protocols in Molecular Biology(JohnWileyandSons,Inc.).

2.Kalintsky,G.,Hummel,J.P.andDierks,C.(1959)J.Biol.Chem.234,1512-1516.

RNase I

Molecular Biology Enzymes | Nucleases—RNases

78020Y 100 mgCustoms by request


Description:RNaseAspecificallycleavessingle-strandedRNAat3'phosphatelinkagesofpyrimidineresiduesleavingpyrimidine3'phosphatesandoligonucleotideswithterminalpyrimidine3'phosphates(1).

Applications:�� CleavingunhybridizedareasofRNAfromRNA:DNAhybridsinRNAorDNAmapping(2)

�� RemovingcontaminatingRNAfromDNAmini-preps.(Add1µlofa10mg/mlRNaseAsolutiontoresuspendedDNApelletpreparedfroma1.5mlbacterialculture)


Properties:MolecularWeight:13.7kDaInhibitors:Ribonucleaseinhibitor;vanadyl-

ribonucleosidecomplexes

Purity:TheenzymeischromatographicallypurifiedandistestedforcontaminatingproteasesandDNases.

Assay conditions:TwounitsofRNaseAareincubatedwithTorulayeastRNAin0.05Msodiumacetate,pH5.0,inavolumeof3mlat25°C.Absorbanceat300nmisreadat15secondintervalsfor4minutes;incubationthencon-tinuesat25°C.Absorbanceistakenat300nmagainafter3.5hoursoruntilasteadystateisobtained.

Unit definition:OneunitistheamountofenzymerequiredtocatalyzethehydrolysisofRNAataratesuchthatK(velocityconstant)equalsunity(Kunitzunits)atpH5.0and25°C.

Concentration:Approximately80units/mg


References:1.Davidson,J.N.(1972) The Biochemistry of the

Nucleic Acids,7thedition,(AcademicPress,NewYork).

2.Myers,R.M.,Larin,Z.andManiatis,T.(1985)Science230,1242-1246.

RNase A, Lyophilized (E.C.3.1.27.5)

27-0323-01 100 mg

Source: Bovinepancreas

Description: RNaseI‘A’specificallycleavessingle-strandedRNAat3’phosphatelinkagesofpyrimidineresiduesleav-ingpyrimidine3’phosphatesandoligonucleotideswithterminalpyrimidine3’phosphates(1).

Applications: �� CleavingunhybridizedareasofRNAfromRNA:DNAhybridsinRNAorDNAmapping(2).�� RemovingcontaminatingRNAfromDNAmini-preps.(Add1μlofa10mg/mlRNaseAsolutiontoresuspendedDNApelletpreparedfroma1.5mlbacterialculture.)

Form: Lyophilizedpowder

Properties: MolecularWeight:13.7kDa(RNaseA),14±0.3

kDa(RNaseB)pI:9.45OptimumpH:7.0-7.5Inhibitors:Heavymetalions,RNaseinhibitors,

vanadyl-ribonucleosidecomplexes.Inactivation: InactivationofRNaseIcanbespecifi-callyachievedusingaRNaseinhibitor.RemovalofRNaseinhibitoristypicallyachievedbytreatmentwithProteinaseKfollowedbyphenolextractionsandethanolprecipitations.

Purity: RNaseIcontains70%RNaseAandtheremaining30%isotherisozymesofRNase.ThisRNaseIisasaltfree,chromatographicallypreparedfreezedriedpowder. Testedforproteaseactivity.

Unit definition: OneKunitzunitcatalyzesthehydrolysisofyeast

RNAtoyieldafirst-ordervelocityconstantof1.0at25°C,pH5.0.

Concentration: ≥70Kunitzunits/mg

Notes: Activeunderawiderangeofconditions.Underlow-saltconditions(0-100mMNaCl),single-strandedRNA,double-strandedRNA,andtheRNAstrandofanRNA:DNAduplexarecleaved.Withhighsalt(>300mMNaCl),single-strandedRNAiscleaved.

Shipping and storage: Shippedondryice.Storeat-20°C.

References:1.Davidson,J.N.(1972)The Biochemistry of the


2.Myers,R.M.,Larin,Z.,andManiatis,T.(1985)Science,230,1242-1246.

RNase I ‘A’, Powder


Molecular Biology Enzymes | Nucleases—RNases

70194Y 1 mg70194Z 5 mgCustoms by request


Description:RNaseAspecificallycleavessingle-strandedRNAat3'phosphatelinkagesofpyrimidineresiduesleavingpyrimidine3'phosphatesandoligonucleotideswithterminalpyrimidine3'phosphates(1).Thisenzymedoesnotrequireco-factorsanddivalentcationsfortheactivity.

Applications:�� CleavingunhybridizedareasofRNAfromRNA:DNAhybridsinRNAorDNAmapping(2)

�� RemovingcontaminatingRNAfromDNAmini-preps.(Add30unitsRNaseAsolutiontoresuspendedDNApelletpreparedfroma1.5mlbacterialculture)�� Preparationofrecombinantproteins�� Ribonucleaseprotectionassays(3)

�� PlasmidandgenomicDNAisolation(4,5)

�� Mappingsingle-basemutationsinDNAorRNA(6,7)

Properties:MolecularWeight:13.7kDaInhibitors:Ribonucleaseinhibitor;vanadyl-

riboncleosidecomplexes;heavymetalionsOptimumpH:7.0-7.5

Purity:TheenzymeischromatographicallypurifiedandistestedforproteaseandDNaseactivities.Itisnotnecessarytoboiltheenzymebeforeuse.

Storage buffer:50mMsodiumacetate,(pH5.0)0.3mMEDTA,50%glycerol.

Unit definition:OneunitofRNaseAistheamountofenzymerequiredtocauseanincreaseinabsorbanceof1.0at260nmat37°C(pH5.0)whenyeastrRNAishydrolyzedtoacid-solubleoligonucleotides.OneKunitzunitisequivalentto50USBunits.

Concentration:≥ 20units/µl;~5mg/ml

Shipping and storage:Shippedondryice.Storeat4°Cor-20°C.

References:1.Davidson,J.N.(1972) The Biochemistry of the


2.Myers,R.M.,Larin,Z.andManiatis,T.(1985)Science230,1242-1246.

3.Sambrook,J.,Russell,D.W.(2001)Molecular Cloning: A Laboratory Manual,thethirdedition,ColdSpringHarborLaboratoryPress,ColdSpringHarbor,NewYork,7.63-7.74.

4.Sambrook,J.,Russell,D.W.(2001)Molecular Cloning: A Laboratory Manual,thethirdedition,ColdSpringHarborLaboratoryPress,ColdSpringHarbor,NewYork,1.31-1.38.

5.Sharma,R.C.,MurphyA.J.,DeWald,M.G.andSchimke,R.T.(1993)BioTechniques14,176-178.

6.MyersR.M.,Larin,Z.andManiatis,T.(1985)Science230,1242-1246.

7.WinterE.,Yamamoto,F.,Almoguera,C.andPerucho,M.(1985)Proc. Natl. Acad. Sci.USA,82,7575-7579.

RNase A, Molecular Biology Grade, Solution (E.C.3.1.27.5)


Source:E. colistraincontaininganoverproducingcloneofE. coliRNaseH.

Description:E. coliRNaseHisanendoribonucleasethatdegradestheRNAportionofDNA:RNAhybrids(1).ItisakeyenzymeintheOkayama-BergandGubler-HoffmancDNAsynthesisandcloningprocedures,inwhichitisemployedtoremovethemRNAduringsecond-strandcDNAsynthesis(2,3).RNaseHisusefulfortheremovalofpoly(A)tailsonmRNAsafterhy-bridizationwitholigo(dT)(4-6).IthasalsobeenusedforthespecificfragmentationofRNAafterhybridiza-tionoftheRNAwitholigodeoxynucleotides(7).

Applications:�� RemovesthemRNAstrandoftheRNA:DNAduplexduringsecondstrandcDNAsynthesis�� Removalofpoly(A)tailsonmRNAafterhybridizationwitholigodT�� MakingspecificfragmentationofRNAafterhybridizationofRNAwitholigodeoxynucleotides�� Site-specificcleavageofRNA(7)

�� Studyofin vitropolyadenylationreactionproducts(8)

Properties:Activators:Mg2+orMn2+

MolecularWeight:21kDa

Purity:Testedforcontaminatingnon-specificendonucle-ases,exonucleases,andribonucleases.

Storage buffer:20mMTris-HCl(pH7.9),100mMKCl,10mMMgCl2,0.1mMEDTA,0.1mMDTT,50%glycerol.

Assay conditions:Reaction(100µl)contains20mMTrisHCl(pH8.0),100mMKCl,10mMMgCl2,1mMDTT,0.5nmolpoly(rA):poly(dT),enzyme.

Unit definition:Oneunitcatalyzesthehydrolysisof1nmolofRNAinradiolabeledpoly(A)-poly(dT)toacid-solublematerialin20minutesat37°C.


5X RNase H Reaction Buffer (1 ml included):100mMTris-HCl(pH7.5),100mMKCl,50mMMgCl2,0.5mMEDTA,0.5mMDTT


References:1.Crouch,R.J.(1981)in Gene Amplification and

Analysis,eds.J.G.ChirikjianandT.S.Papas,(Elsevier/North Holland, Inc., New York)2,217-228.

2.Okayama,H.andBerg,P.(1982)Mol.Cell Biol.2,161-170.

3.Gubler,U.andHoffman,B.J.(1983)Gene 25,263-269.

4.Vournakis,J.N.,Efstratiadis,A.andKafatos,F.C.(1975)Proc.Natl.Acad.Sci.USA72,2959-2963.

5.Narayan,P.,Ayers,D.F.,Rottman,F.M.,Maroney,P.A.andNilsen,T.W.(1987)Mol.Cell.Biol.7, 1572-1575.

6.Davis,R.E.,Davis,A.H.,Carroll,S.M.,Rajkovic,A.andRottman,F.M.(1988)Mol.Cell Biol.8,4745-4755.

7.Donis-Keller,H.(1979)Nucl.Acids Res.7,179-192.

8.Goodwin,E.C.andRottman,F.M.(1992)Nucl. Acids Res.20,916.

RNase H


Molecular Biology Enzymes | Nucleases—Non-Specific

70196Y 15,000 unitsCustoms by request

Source:Staphylococcus aureus

Description:MicrococcalNucleaseisasingle-anddouble-strand-edDNAandRNAendonucleasewhichyields3'nucleotides(1).Itismorespecificforsingle-strandednucleicacidsandshowspreferenceforAUorAT-richregions.Completedigestionyieldsmono-anddinucleotides.

Applications:�� RemovalofRNAandDNAfromproteinpreparations�� RemovalofmRNAinareticulocytelysate�� Analysisofchromatinstructure


Properties:IsoelectricPoint:9.6OptimumpH:8.8-9.2OptimumTemperature:37°CMolecularWeight:16.81kDa(2)

RequirementforDivalentCation:Ca2+foractivityInhibitors:EDTAorEGTA

Purity:Testedforcontaminatingproteases.

Solubility and storage conditions:1.Shorttermstorage:Dissolve1mg/mlMicrococcal

NucleaseindH2O.Storeat4°Cfor1-2weeks.2.Longtermstorage:DissolveindH2Ocontaining

20-50%glycerolat1mg/mlconcentration.Storeat-20°C.Donotfreezeandthawmorethanonetime.

Assay conditions: Thereactionmixturecontains28mMsodiumborate,pH8.0,1.4mMCaCl2,100µgDNA,andenzyme.Incubationisat37°Cfor30minutes.

Unit definition:OneunitistheamountofenzymewhichchangestheopticaldensityofaDNAsubstrateby1.0at260nmatpH8.0,37°C(3).

Concentration:≥5,000units/mg

Shipping and storage:Shippedondryice.Seepreviousinformation.

References:1.Alexander,M.,Heppel,L.A.andHurwitz,J.(1961)

J.Biol.Chem.236,3014-3019.2.Taniuchi,H.,Anfinsen,C.B.andSodja,A.(1967)

J.Biol.Chem.242,4752-4758.3.Heins,J.N.,Taniuchi,H.andAnfinsen,C.B.

(1966)Procedures in Nucleic Acid Research,eds.G.L.CantoniandD.R.Davies(HarperandRowPublishers,Inc.,NewYork),79.

4.Cone,J.L.,Cusumano,C.L.,Taniuchi,H.andAnfinsen,C.B.(1971)J.Biol.Chem.246,3103-3110.

5.Ausubel,F.,Brent,R.,Kingston,R.E.,Moore,D.D.,Seidman,J.G.,Smith,J.A.andStruhl,K.(1989)Current Protocols in Molecular Biology.(JohnWileyandSons).

Micrococcal Nuclease (E.C.3.1.31.1)

20240Y 100 unitsCustoms by request

Source:Crotalus adamanteusvenom

Description:PhosphodiesteraseI(venomexonuclease)iswidelyusedinstudyingnucleicacidstructureandsequence(1).Ithydrolyzes5'-mononucleotidesfrom3'-hydroxy-terminatedDNAandRNA(2).Phosphodi-esteraseIcleavesADP-ribosylatedproteinsatthepy-rophosphatelinkagestoyieldphosphoribosyl-AMP(3).

Application:�� Basecompositionand“nearestneighbor”analysis(1)


Properties:MolecularWeight:115kDaOptimumpH:9.8-10.4Activators:Mg2+

Inhibitors:5mMEDTA,reducingagents(gluthathione,cysteine,andascorbicacid),partialinhibitionbyATP,ADP,andAMP

Purity:PartiallypurifiedbythemethodofBjörk(4)andtreatedtoremovecontaminating5'nucleotidase(5).

Assay conditions: Thereactionmixturecontains100mMTris-HCl,pH8.9,100mMNaCl,14mMMgCl2,0.5mMp-nitrophenylthymidine-5'-phosphate,andPhospho-diesteraseI.

Unit definition:Oneunithydrolyzes1µmolp-nitrophenylthymidine-5-phosphateperminutesat25°C,pH8.9.

Concentration:≥20units/mg

Resuspension and Storage: Dissolveat1mg/mlin110mMTris-HCl,pH8.9,110mMNaCl,15mMMgCl2,and50%glycerol.Storeat-20°Cafterrehydration.


References:1.Ho,N.W.andGilham,P.T.(1973)Biochem.

Biophys.Acta,308,53-58.2.Laskowski,M.(1971)inThe Enzymes, 3rd Ed.

(Academic Press, New York)IV,313-328.3.Hayaishi,O.(1976)Trends Biochem.Sci.1,9.4.Björk,W.,(1963)J. Biol. Chem.238,2487.5.Sulkowski,E.andLaskowski,M.(1971)Biochem.

Biophys.Acta,240,443.

Phosphodiesterase I(E.C.3.1.4.1)


Molecular Biology Enzymes | Phosphatases—Alkaline Phosphatases

70034Y 1,500 unitsCustoms by request

Source:Bovineintestinalmucosa

Description:CalfIntestinalAlkalinePhosphatase(CIAP)catalyzesthehydrolysisofmanyphosphomonoesters,includ-ing5'-nucleotides,RNA,andDNA.Itiswidelyusedforthedephosphorylationof5'-phosphorylatedendsofDNAorRNAforsubsequentlabelingwith32Pusing[g-32P]ATPandT4PolynucleotideKinase(PN70031).Dephosphorylationalsopreventsreliga-tionoflinearizedplasmidDNAincloningexperi-ments.CIAPcanbeinactivatedbyphenol:chloroformextraction,bygelpurification,orbyspincolumn.

Applications:�� Dephosphorylationof5'terminiofnucleicacidsbeforelabelingwithT4PolynucleotideKinase�� Dephosphorylationof5'terminiofDNAfragmentstopreventself-ligation

Properties:MolecularWeight:Dimerofapproximately110kDa

(two55kDasubunits)Activators:Zn2+,Mg2+,Ca2+

IsoelectricPoint:5.7RequirementforStability:TritonX-100,zincacetate

orzincchloride,cysteineOptimumpH:9-10OptimumTemperature:37°C.pHStability:pH7.5;5.0inthepresenceofTriton

X-100,Zn2+,cysteine

Purity:Testedforcontaminatingnon-specificendonucle-ases,exonucleases,andribonucleases.

Storage buffer:10mMTris-HCl(pH7.5),5mMMgCl2,0.1mMZnCl2,50%glycerol.

Assay conditions:Thereactionmixturecontains1Mdiethanolamine,pH9.8,0.25mMMgCl2,10mMp-nitrophenylphosphate,and0.01-0.1unitsofCalfIntestinalAlkalinePhosphatase(CIAP).Reactiontemperatureis37°C.Thechangeinabsorbanceat405nmismonitored(3050µlreactionvolume).

Unit definition:Oneunitofenzymecatalyzesthehydrolysisof1µmolofp-nitrophenylphosphateperminuteatpH9.8indiethanolaminebufferat37°C.


Functional test:Dephosphorylationofarestrictionenzymedigestedplasmid(5-10pmolof5'ends,0.01-0.05units/pmol5'ends.Reducesreligationto<0.5%com-paredtotheuntreatedcontrol.)

Functionally tested 10X CIAP Reaction Buffer (1 ml included):200mMTris-HCl(pH8.0),100mMMgCl2.

CIAP Dilution Buffer, 10X (1 ml included):50mMTris-HCl(pH8.0).


References:1.Zhang,Y.,Zhu,Y.andZhou,H.(2000)Int. J.

Biochem. Cell. Biol.32,887-894.2.Abdolrazaghi,Z.andButterworth,P.(1983)

Enzyme30,12-20.3.Baunoch,D.A.,et al.(1992)Oncogene,7,2351-

2353.

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T4 Polynucleotide Kinase70031Y 500 units70031Z 1,000 units70031X 2,500 units

Calf Intestinal Alkaline Phosphatase (CIAP) (E.C.3.1.3.1.)


78390 500 units1,000 units

5,000 units

Proven performance – the phosphatase benchmark�� Purifiedfromarecombinantsource�� 100%heat-inactivatedin5minutesat65°C�� Significantlyimprovedstoragestabilityatlowertemperatures(seeFig.1)�� Veryhighspecificactivity�� Removes5’-phosphatesfromDNA,RNA,dNTPs,andproteins�� Maybeaddeddirectlytorestrictionenzymedigests�� Novectorpurificationnecessary�� Requiresnosupplementalzincorotheradditivesforactivity�� Worksdirectinmanydifferentbuffers�� EasytreatmentofunincorporateddNTPsinPCRproductspriortoDNAsequencingorSNPanalysis

Source: Recombinant

Description:ShrimpAlkalinePhosphatase(SAP)isahighspecificactivity,heat-labilealkalinephosphatasepurifiedfromarecombinantsourceandoriginallyisolatedfromPandalus borealis(arcticshrimp).SAPisusefulinmanymolecularbiologyapplicationssuchasthedephosphorylationofphosphorylatedendsofDNAorRNAforsubsequentuseincloningorend-label-ingofprobes.Incloning,dephosphorylationpreventsreligationoflinearizedplasmidDNA.SAPmayalsobeusedtotreatunincorporateddNTPsinPCRreac-tionstopreparetemplatesforDNAsequencingorSNPanalysis.

SAPhasapproximatelythesamespecificactivityasCalfIntestinalAlkalinePhosphatase(CIAP),andlikeCIAP,isactiveinvirtuallyallrestrictionenzymereactionbuffers.UnlikeCIAP,SAPiscompletelyandirreversiblyinactivatedbyheatingreactionsat65°Cfor15minutes.

Applications:�� DephosphorylationofDNApriortocloning�� PCRcleanupforsequencingandSNPapplications�� DephosphorylationofDNApriortoend-labellingusingT4PNKorOptiKinase�� DephosphorylationofRNA�� Proteindephosphorylation

Properties:MolecularWeight:Homodimer.Monomeris55kDa

asdeterminedbyaminoacidsequence.OptimumpH:10.4inglycinebufferandpH8.0in

Trisbuffer.OptimumTemperature:37°CHeat-Inactivation:65°Cfor15minutesInhibitors:10mMDTT,0.1%β-MEReactionConditions:ActiveinNaClandKCl.

RequiresMg2+forhighestactivity.

Purity:Testedforcontaminatingendonucleases,exonucle-ases,andribonucleases.

Storage buffer:25mMTris-HCl(pH7.5),1mMMgCl2,50%glycerol.

Assay conditions:Thereactionmixturecontains100mMglycine,pH10.4,1mMMgCl2,1mMZnCl2,10mMp-nitrophenylphosphate,and0.001–0.1unitsofShrimpAlkalinePhosphatase(SAP).Thechangeinabsorbanceat405nmismonitored(3050μlreac-tionvolume).

Unit definition:Oneunitistheamountofenzymewhichcatalyzesthehydrolysisof1μmolofp-nitrophenylphosphateperminuteinglycinebuffer(pH10.4)at37°C.

Concentration:1unit/μl

Functional test:Dephosphorylationofrestrictionenzymedigestedplasmids(5–20pmolof5’-ends,0.1–0.5units/pmol5’-ends).Reducesreligationto<0.5%com-paredtotheuntreatedcontrol.

Functionally tested 10X SAP Reaction Buffer (1 ml included):200mMTris-HCl(pH8.0),100mMMgCl2.

Functionally tested SAP Dilution Buffer (1 ml included):50mMTris-HCl(pH8.0).

References:1.Ruan,C.C.,Samols,S.B.andFuller,C.W.(1990)

Comments17,(No.1),UnitedStatesBiochemicalCorporation,Cleveland,OH.

2.Werle,E.,ScneiderC.,Renner,M.,Völker,M.andFiehn,W.(1994)Nucl. Acids Res.22,4354-4355.

3.Hanke,M.andWink,M.(1994)BioTechniques17,858-860.

Shrimp Alkaline Phosphatase (SAP)


Fig. 1. Exceptional stability of SAP. ThreemonthstabilityofSAPatvarioustemperatures.SAPwasstoredfor3monthsattheindicat-edtemperaturesandtheremainingactivitywasmeasuredusingastandardp-nitrophenylphosphataseactivity.

0%

20%

40%

60%

80%

100%

-20ºC 4ºC 25ºC 37ºC

Rem

aini

ng A

ctiv

ity

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PCR Product Pre-Sequencing Kit70995 100 reactions70996 500 reactions70997 2,000 reactions



75592 50 reactions

Source:RecombinantPandalus borealis(arcticshrimp)

Description:SuperSAPisanenhancedformulationofShrimpAlkalinePhosphatasedesignedforrapiddephos-phorylationofDNApriortoitsuseincloningapplicationsandend-labeling.SuperSAPdephos-phorylatesupto5pmolsofcohesiveorbluntendswithabackgroundreduction≥95%injust5minutes.Backgroundreductionistheinhibitionofrecircularizationinaself-ligationreactionandmeasuredbytransformationinE. coli.SuperSAPmayalsobeusedtodephosphorylateRNAandprotein.

BenefitsSuperSAPhasallthebenefitsofSAP.Ithasahighspecificactivityandisheat-labile.SuperSAPcanbeeasilyheat-inactivatedat65°Cfor15minutesandworkswellinvariousrestrictionenzymebuffers.Noadditionalbuffersoradditivesarerequired.

Rapid dephosphorylationSuperSAPremovestheterminal5'phosphatefromprotruding-,blunt-orrecessed-endsinjust5minutes(Table1).

Easy vector preparation for cloningSuperSAPworksdirectlyinmostrestrictionenzymedigestsandcanbeaddeddirectlytothereactiontosimultaneouslycutanddephosphorylateDNA.Noadditionalbuffersoradditivesarerequired(Table2).

Convenient – Direct ligationSuperSAPreactionsdonotrequirepurification,e.g.phenolextraction,ethanolprecipitation,orspincol-umn,priortoligation.AfterSuperSAPtreatment,thedephosphorylatedcutvectormaybedirectlyligatedusingtheLigate-ITRapidLigationKit[PN78400/10]orT4DNALigase[PN70005].

Purity:Testedforcontaminatingendonucleases,exonucle-ases,andribonucleases.

Storage buffer:25mMTris-HCl,pH7.6,1.0mMMgCl2,0.1mMZnCl2,and50%glycerol.

10X SuperSAP Reaction Buffer (1 ml included): 200mMTris-HCl,pH8.0,100mMMgCl2.

Functional test:SuperSAPhasbeenfunctionallytestedinthefollow-ingprotocols:

Functional test: 5 Minute DephosphorylationDephosphorylationof5pmolofpurifiedvectorDNAterminiwith1µlSuperSAPin1XReactionBufferin5minutesat37°Creducesthebackgroundofreligationandtransformationto>95%comparedtountreatedcontrol.

Simultaneous dephosphorylation and restric-tion enzyme digestionSimultaneousdephosphorylationandrestrictionenzymedigestwith1µlofSuperSAPfor5'overhangandbluntendsand2µlfor3'overhangin30min-utesat37°Creducesthebackgroundofreligationandtransformationto>95%comparedtountreatedcontrol.

Components: 50Reaction PackSizeSuperSAP 50µl10XSuperSAPReactionBuffer 1ml


References:1.Ruan,C.C.,Samols,S.B.,andFuller,C.W.(1990)

Comments17,(No.1),UnitedStatesBiochemicalCorporation,Cleveland,OH.

2.Werle,E.,Scneider,C.,Renner,M.,Voelker,M.andFiehn,W.(1994)Nucl. Acids Res.22,4354-4355.

3.Hanke,M.andWink,M.(1994)BioTechniques17,858-860.

Table 1

Number of Background colonies reduction (%)

5' OverhangEcoR I-cut pUC 19 vectorControl 5831µlSuperSAP 5 99%

3' OverhangPst I-cut pUC 19 vectorControl 19871µlSuperSAP 0 100%

Blunt EndHinc II-cut pUC 19 vector Control 4901µlSuperSAP 2 99.6%

Rapid SuperSAP dephosphorylation for 5 minutes prevents recircularization of vector. pUC19vectorwasdigestedasindicat-ed,purifiedusingaspincolumnandre-suspendedin10mMTris-HCl,pH8.5.5µgwastreatedwith1µlofSuperSAPandincubatedfor5minutesat37°Cfollowedbyheat-inactivationat65°Cfor15min-utes.50ngwasself-ligateddirectlyusingtheLigate-ITRapidLigationKit(PN78400),2.5ngwastransformedintoE. coliDH5-aand0.5ngwasplatedonselectivemedium.

Table 2

Number of Background colonies reduction (%)

Kpn I-cut pUC 19 vector Buffer L (USB) 3' Overhang Control 47962µlSuperSAP 44 99%

Hind III-cut pUC 19 vector Buffer M (USB) 5' Overhang Control 17831µlSuperSAP 13 99.3%

EcoR I-cut pUC 19 vector Buffer H (USB) 5' Overhang Control 32691µlSuperSAP 84 97.4%

BamH I-cut pUC 19 vector Buffer K (USB) 5' Overhang Control 17531µlSuperSAP 0 100%

Sma I-cut pUC 19 vector Buffer A (USB) Blunt End Control 10911µlSuperSAP 0 100%

Simultaneous dephosphorylation with SuperSAP and restriction enzyme digestion is convenient and effective to prevent recircularization of vector. 1µgofpUC19wasdigestedasindicatedwith1µlofSuperSAP(5'overhangorblunt),or2µlofSuperSAP(3'overhang)intheappropriaterestrictionenzymebufferandwasincubatedfor30minutesat37°Cfollowedbyheat-inactiva-tionat65°Cfor15minutes.50ngwasdirectlyself-ligatedusingtheLigate-ITRapidLigationKit(PN78400),2.5ngwastransformedintoE. coliDH5-aand0.5ngwasplatedonselectivemedium.

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SuperSAP Shrimp Alkaline Phosphatase


Molecular Biology Enzymes | Phosphatases—Pyrophosphatase

78450 5 units 50 unitsCustoms by request

Source:PurifiedfromE. colicellsexpressingtheclonedppagene(1)fromSaccharomyces cerevisiae(Baker’sYeast).

Description:DNAandRNApolymerasereactions,aswellasotherenzymaticreactions,generateinorganicpyrophos-phateasaby-product.Thisby-productinhibitsnucleicacidsynthesisthroughaprocesscalledpy-rophosphorolysiswhichreversesthepolymerizationreactionandlimitsDNA/RNAsynthesis.InorganicpyrophosphataseispartoftheDNA/RNAsynthesispathwayin vivo,renderingpolymerizationessentiallyirreversiblebyremovingpyrophosphate(2).Inorganicpyrophosphatase(PPase)catalyzesthehydrolysisofinorganicpyrophosphatetoformtwomoleculesoforthophosphate(3):P2O7

-4+H2O→2HPO4-2

Inorganicpyrophosphatasemaybeusedtoremovepyrophosphateandimprovetheresultsoftechniquesthatareparticularlysensitivetoitsaccumulationsuchassingle-baseextension(SBE)(4)andSangersequencing,especiallywithregardtoselectivebandweakening(5,6).Also,inorganicpyrophosphatasecangreatlyincreasetheyieldofin vitrotranscriptionreactions(7).

Applications:�� PreventsaccumulationofpyrophosphateduringDNAsequencing.�� High-yieldsynthesisofRNAbyin vitrotranscription.�� RemovalofcontaminantPPipriortoSNPgenotypingbasedonthedetectionofpyrophosphate.�� RemovalofcontaminantPPipriortosingle-base-extension(SBE).

Properties:MolecularWeight:70kDahomodimer(8)

(2x35kDa)pI:4.75Activators:Mg+2>Zn+2>Co+2>Mn+2>Ca+2,note

thathydrolysisofinorganicpyrophosphateisspecificinthepresenceofMg+2butbothADPandATPcanbehydrolyzedifzincispresent(9).

Inhibitors:EDTAInactivation:Inactivatedbyphenol/chloroform

extractionIVT:RecommendedconcentrationofrPPaseduringin

vitrotranscriptionreactionsis1-10units/ml

Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingexonucleases,endonucle-ases,andribonucleases.

Storage buffer:10mMTris-HCl(pH7.5),0.1mMEDTA,50%glycerol.

rPPase Dilution Buffer (1 ml included):10mMTris-HCl(pH7.5),0.1mMEDTA,50%glycerol.

Unit definition:Oneunithydrolyzes1µmolofpyrophosphatein1minuteat25°C.

Concentration:0.1units/µl

Functional test:DNAsequencingusingtheSequenaseVersion2.0DNASequencingKit.

Shipping and storage:Shippedondryice.Storeat-20°C

References:1.Kurilova,S.A.,Vorobjeva,N.N.,Nazarova,T.I.

andAvaeva,S.M.(1993)FEBS Letters333(3),280-282.

2.Kornberg,A.(1980)DNAReplication(W.H.Freeman&Co.SanFrancisco),54-55.

3.Cooperman,B.S.(1982)Methods in Enzymology87,526-548.

4.Zhou,G.H.,Shirakura,H.,Kamahori,M.,OkanoK.,NagaiK.andKambara,H.(2004)Human Mutation24(2),155-163.

5.Tabor,S.andRichardson,C.C.(1990)J. Biol. Chem.265,8322-8328.

6.Ruan,C.C.,Samols,S.B.andFuller,C.W.(1990)Comments17,No.2,UnitedStatesBiochemicalCorporation,Cleveland,OH.

7.Cunningham,P.R.andOfengand,J.(1990)BioTechniques9,713-714.

8.Butler,L.G.(1971)Enzymes(AcademicPressNewYork),529-540.

9.Knight,W.B.,Dunaway-Mariano,D.,Ransom,S.C.andVillafranca,J.J.(1984)J. Biol. Chem.259(5),2886-2895.

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Pyrophosphatase, Inorganic (Recombinant) (rPPase)


70057Y 125 units70057Z 750 units70057 2,500 unitsCustoms by request

Source:E. coli straincontaininganoverproducingcloneofE. coli Exonuclease-freeKlenowfragment.

Description:ThelargefragmentofDNApolymeraseIfromE. coli,Klenow,hasbeengeneticallyengineeredtoeliminatethe3'→5'exonucleaseactivitynormallyassociatedwiththispolymerase(1).Exo-freeKlenowisusefulinapplicationswhereaccompanyingexonucleaseactivityisofsignificantconcern,suchasrandomprimedlabeling,DNAsequencingbytheSanger,et al.method,orstranddisplacementamplification(SDA).

Applications:�� RandomprimedDNAlabeling�� Stranddisplacementamplification(SDA)(2)

�� DNAsequencingbytheSangerdideoxymethod(3)

�� Labelingrecessed3'terminiofDNAfragmentswithlabeleddNTP

Note:Exonuclease-FreeKlenowisnotrecommendedforfill-inreactionsbeforeDNAligation,sinceitfrequentlyaddsoneormoreextranucleotidestothe3'-terminusofablunt-endedDNAsubstrateinanon-templatedirectedfashion(4).

Properties:MolecularWeight:68kDa

Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingdouble-strandedendonu-cleasesandexonucleases.

Storage buffer:50mMpotassiumphosphate(pH7.0),1.0mMDTT,1.0mMEDTA,50%glycerol.

Assay conditions: Thereactionmixturecontains100mMpotassiumphosphate(pH7.4),6.7mMMgCl2,1mMDTT,100µMeachdNTP,radiolabeleddATP,400µg/mlactivatedsalmonspermDNA,andenzyme.Incuba-tionisfor30minutesat37°C.

Unit definition:Oneunitistheamountofenzymerequiredtocatalyzetheincorporationof10nmoloftotaldeoxy-ribonucleotidesintoanacid-precipitablematerialin30minutesat37°C.


Functional test:Filling-in3'recessedendswith>50%incorporationofradiolabeleddATPinto4µgofrestrictionenzymedigestedDNAin15minutesat30°C.

Functionally tested 10X Exonuclease-Free Klenow Reaction Buffer (1 ml included):0.5MTris-HCl(pH7.4),0.1MMgCl2,10mMDTT.


References:1.Derbyshire,V.,Freemont,P.S.,Sanderson,M.R.,

Beese,L.,Friedman,J.M.,Joyce,C.M.andSteitz,T.A.(1988)Science 240,199-201.

2.Walker,G.T.(1993)PCR Methods Appl.3,1-6.3.Sanger,F.,Nicklen,S.andCoulsen,A.R.(1977)

Proc. Natl. Acad. Sci. U.S.A. 74,5463-5467.4.Clark,J.M.,Joyce,C.M.andBeardsley,G.P.

(1987)J. Mol. Biol.198,123-127.

Exonuclease-Free Klenow (LargeFragmentofDNAPolymeraseI,Exonuclease-Free,E.C.2.7.7.7)

Molecular Biology Enzymes | Polymerases—DNA Polymerases

72020 750 units2,500 units

Source:E. colistraincontaininganoverproducingcloneofE. coliExonuclease-freeKlenowfragment.

Description:ThelargefragmentofDNApolymeraseIfromE. coli,Klenow,hasbeengeneticallyengineeredtoeliminatethe3’→5’exonucleaseactivitynormallyassociatedwiththispolymerase(1).Exo-freeKlenowisusefulinapplicationswhereaccompanyingexonucleaseactivityisofsignificantconcern,suchasrandomprimedlabeling,DNAsequencingbytheSanger,et al.method,stranddisplacementamplifica-tion(SDA),orwholegenomeamplification(WGA).Exonuclease-FreeKlenow,TrisBufferisstoredinaphosphatefreebuffer

Applications:�� RandomprimedDNAlabeling�� Stranddisplacementamplification(SDA)(2)

�� DNAsequencingbytheSangerdideoxymethod(3)

�� Labelingrecessed3'terminiofDNAfragmentswithlabeleddNTP

Note:Exonuclease-FreeKlenowisnotrecommendedforfill-inreactionsbeforeDNAligation,sinceitfrequentlyaddsoneormoreextranucleotidestothe3'-terminusofablunt-endedDNAsubstrateinanon-templatedirectedfashion(4).


Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingdouble-andsingle-strand-edendonucleasesandexonucleases.

Storage buffer:25mMTris-HCl,pH7.5,1mMDTT,0.1mMEDTAand50%glycerol

Assay conditions: Thereactionmixturecontains100mMpotassiumphosphate(pH7.4),6.7mMMgCl2,1mMDTT,20μMpoly(dA-dT)-(dA-dT),33μMdATP,33μMdTTP,andenzyme.Incubationisfor30minutesat37°C.



Functional test:Filling-in3’recessedendswith>50%incorporationofradiolabeleddATPinto0.1-4μgofrestrictedDNAin15minutesat30°C.

Functionally Tested 10X Exonuclease-Free Klenow Reaction Buffer (1 ml included):0.5MTris-HCl(pH7.4),0.1MMgCl2,10mMDTT.

References:1.Derbyshire,V.,Freemont,P.S.,Sanderson,M.R.,

Beese,L.,Friedman,J.M.,Joyce,C.M.andSteitz,T.A.(1988)Science 240,199-201.

2.Walker,G.T.(1993)PCR Methods Appl.3,1-6.3.Sanger,F.,Nicklen,S.andCoulsen,A.R.(1977)

Proc. Natl. Acad. Sci. U.S.A. 74,5463-5467.4.Clark,J.M.,Joyce,C.M.andBeardsley,G.P.

(1987)J. Mol. Biol.198,123-127.

Exonuclease-Free Klenow, Tris Buffer(LargeFragmentofDNAPolymeraseI,Exonuclease-Free)




Source:E. coli straincontaininganoverproducingcloneofE. coli Klenowfragment.

Description:Klenowenzyme,thelargefragmentofDNAPolymeraseIfromE. coli,retainspolymerizationand3'→5'exonucleaseactivitybutlacksthe5'→3'exonucleaseactivity.Itiswell-suitedfortheconversionof5'-protrudingendstoflushendsforcloning(1),in3'-endlabelingofDNAfragmentsforsequenceanalysis(2),forsecond-strandsynthesisofcDNA(3),andforrandomprimedlabelingofDNAfragments.

Applications:�� Fillinof3'recessedterminiproducedbysomerestrictionenzymesbeforeligation�� SecondstrandcDNAsynthesis�� Labeling3'terminiofDNAfragmentswithlabeleddNTP

Properties:MolecularWeight:68kDaOptimumpH:7.4inphosphatebuffer;8.4inTris-HClOptimumTemperature:37°CRequirementsforDivalentCation:Mg2+,Mn2+

OptimumMg2+Concentration:7mM

Purity:Greaterthan98%pureasdeterminedbySDS-PAGE.Testedforcontaminatingdouble-andsingle-strandedendonucleases.

Storage buffer:50mMpotassiumphosphate(pH7.0),1.0mMDTT,50%glycerol.

Assay conditions:Thereactionmixturecontains100mMpotassiumphosphate(pH7.4),6.7mMMgCl2,1mMDTT,100µMeachdNTP,radiolabeleddATP,400µg/mlactivatedsalmonspermDNA,andenzyme.Incuba-tionisfor30minutesat37°C.



Functional test:Filling-in3'recessedendswith>50%incorporationofradiolabeleddATPinto4µgofrestrictionenzymedigestedDNAin15minutesat30°C.

Functionally tested 10X Klenow Reaction Buf-fer (1 ml included):0.5MTris-HCl(pH7.5),100mMMgCl2,10mMDTT,0.5mg/mlBSA.


References:1.Telford,J.L.,Kressmann,A.,Koski,R.A.,

Grosschedl,R.,Muller,F.,Clarkson,S.G.andBirnstiel,M.L.(1979)Proc. Natl. Acad. Sci.USA 76,2590-2594.

2.Joyce,C.M.andGrindley,N.D.F.(1983)Proc. Natl. Acad. Sci. USA 80,1830-1834.

3.Rougeon,F.,Kourilsky,P.andMach,B.(1975)Nucl. Acids Res. 2,2365-2378.

Klenow DNA Polymerase I (LargeFragmentofDNAPolymeraseI,E.C.2.7.7.7)


Source:E. coli straincontaininganoverproducingcloneofE. coli DNAPolymeraseI.

Description:DNAPolymeraseIisaDNAdependent5'→3'polymerasewithinherent3'→5'and5'→3'exonucleaseactivity.Thesethreeactivitiestogethermakethisenzymeusefulforin vitrolabelingofDNAbythenicktranslationmethod(1).ThismethodinvolvestheuseofalowlevelofDNaseIwhichhydrolyzesphosphodiesterbondstocreatenicksatinternallocationsinunlabeledDNA.The3'hydroxylgroupsatthenickingsiteprimeDNAsynthesisbyDNAPolymeraseIwhichconcomitantlydegradestheDNA5'ofthenick.Thisprocess(translation)leavesaradiolabeledpatchofDNAwhichextendsfromtheoriginalnicksitetothesiteoftheremainingnewnickorterminusofafragment.

Applications:�� NicktranslationofDNAinmakingprobes�� Second-strandsynthesisofcDNA

Properties:MolecularWeight:109kDaOptimumpH:7.4inphosphatebuffer;8.4inTris-HClbufferOptimumTemperature:37°CRequirementsforDivalentCation:Mg2+,Mn2+

OptimumMg2+Concentration:7mM

Purity:Greaterthan90%pureasdeterminedbySDS-PAGE.Testedforcontaminatingdouble-strandedendonucleases.

Storage buffer:50mMpotassiumphosphate(pH7.0),1.0mMDTT,50%glycerol,1mMEDTA.

Assay conditions:Thereactionmixturecontains100mMpotas-siumphosphate(pH7.4),6.7mMMgCl2,1.0mMDTT,150µMdNTPs,400µg/mlactivatedDNAastemplate-primer,andenzyme.Incubationisat37°Cfor30minutes.

Unit definition:Oneunitistheamountofenzymerequiredtocata-lyzetheconversionof10nmoloftotaldeoxyribonu-cleotidesintoanacid-insolubleformin30minutesat37°CusingactivatedDNAasthetemplate-primer.



Reference:1.Rigby,P.W.,Dieckmann,M.,Rhodes,C.andBerg,

P.(1977)J. Mol. Biol.113,237-257.

DNA Polymerase I (E.C.2.7.7.7)



Source:E. coli straincontainingclonesthatoverproduceT7Gene5ProteinandE. coli Thioredoxin.

Description:SequenaseVersion2.0DNAPolymeraseisageneticallyengineeredformofT7DNApolymerase(1).Unlikethewild-typeenzymeithasvirtuallyno3'→5'exonucleaseactivity.SequenaseVersion2.0ishighlyprocessive,incorporatesnucleotideanalogs(dlTP,thio-dNTPs,dideoxy-NTPs,etc.),isnotimpededbysecondarystructures,andcancarryoutstranddisplacementsynthesis.Itisanexcellentenzymefordideoxy-sequencingandisusefulinotherapplica-tions,especiallywheretheabsenceofassociatedexonucleaseactivityisdesirable.

Applications:�� DNAsequencing�� Secondstrandsynthesisformicroarrayapplications(2)

�� Randomprimedlabelingwithnucleotideanalogs(2)

�� Labelingthe3'terminiofDNAfragmentswith5'protrudingends�� AmplificationofDNA�� Stranddisplacementapplications

Note:SequenaseVersion2.0DNAPolymerasemaynotbeusefulformakingblunt-endfragmentsbecauseitcanleaveaone-baseover-hangatthe3'end.

Properties:MolecularWeight:Consistsoftwosubunits,

modifiedT7gene5protein(76kDa)andE. colithioredoxin(12kDa).

OptimumpH:7.5OptimumTemperature:37°CRequirementsforDivalentCation:Mg2+,Mn2+

Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingdouble-andsingle-strand-edendonucleasesandexonucleases.

Storage buffer:20mMpotassiumphosphate(pH7.4),1mMDTT,0.1mMEDTA,50%glycerol.

Assay conditions:Thereactionmixture(100µl)contains40mMTris-HCl(pH7.5),10mMMgCl2,5mMDTT,0.3mMdNTPs,and5µgM13mp18pre-annealedto5pmolM13universalprimer;incubationisat37°Cfor1minute.

Unit definition:Oneunitofenzymecatalyzestheincorporationof1nmolofnucleotideintoacidinsolubleformin30secondsat37°C.


Functional test:DNAsequencingwiththeSequenaseVersion2.0DNASequencingKit(PN70770).

Functionally tested 5X Sequenase Version 2.0 Reaction Buffer (1 ml included, PN 70702):200mMTris-HCl(pH7.5),100mMMgCl2,250mMNaCl

Sequenase Version 2.0 Dilution Buffer (1 ml included):10mMTris-HCl(pH7.5),5mMDTT,0.1mMEDTA


References:1.Tabor,S.andRichardson,C.C.(1989)J. Biol.

Chem.264,6447-6458.2.Wang,D.,Coscoy,L.,Zylberberg,M.,Avila,P.C.,

Boushey,H.A.,Ganem,D.andDeRisi,J.L.(2002)Proc Natl. Acad. Sci. USA,99,15687-15692.

3.Paris,M.(1992)Comments 18,(No.3),UnitedStatesBiochemicalCorporation,Cleveland,Ohio.

4.Tabor,S.andRichardson,C.C.(1987)Proc. Natl. Acad Sci. USA 84,4767-4771.

5.Tabor,S.andRichardson,C.C.(1989)Proc. Natl. Acad. Sci. USA 86,4076-4080.

Sequenase Version 2.0 DNA Polymerase (E.C.2.7.7.7)


70017Z 250 units70017 1,000 unitsCustoms by request

Source:E. coli straincontainingoverproducingclonesofT7Gene5andE. colithioredoxin.

Description:T7DNAPolymeraseconsistsoftwosubunits,oneencodedbythebacteriophagegene5andtheotherbytheE. coli trxAgenethioredoxin(1,2).Thisenzymeisresponsiblefortherapidreplicationofbacterio-phageT7DNAduringitsinfectioncycle.InadditiontothehighlyprocessiveDNApolymeraseactivity,italsohashighlevelsofbothsingle-anddouble-strandedDNA3'→5'exonucleaseactivities(3,4).Theexonucleasespecificactivitieshavebeenreportedtorangefrom5%to100%ofthepolymerasespecificactivity(3,5).Theexonucleaseactivityappearstoberesponsibleforthehighfidelityofthisenzymeandpreventsstranddisplacementsynthesis(6).

T7DNAPolymeraseisavaluableenzymewherehighexonucleaseactivityisrequiredorwhereabsenceofstrand-displacementactivityisimportant(6).Ithasalsobeenusedsuccessfullyforsite-directedmutagen-esis(7).This enzyme is not suitable for use in DNA sequencing.ItisdistinctfromSequenaseVersion2.0DNAPolymerasewhichisamodifiedformofT7DNAPolymerasewithno3'→5'exonucleaseactivity.

Applications:�� Labeling3'-terminiofDNAfragmentswithprotruding5'ends(Fill-InReaction)�� Labelingthe3'-terminiofblunt-endedDNAfragmentsortheterminiofDNAfragmentswithprotruding3'-termini�� Site-directedmutagenesis�� NotsuitableforDNAsequencingduetohighexogenouslevelsofexonucleaseactivities

Properties:T7Gene5product(84kDa)andE. colithioredoxin(12kDa).

Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleases.

Storage buffer:20mMpotassiumphosphate(pH7.4),1mMDTT,0.1mMEDTA,50%glycerol.

Assay conditions:Thereactionmixturecontains88mMpotassiumphosphate(pH7.5),6.7mMMgCl2,5mM2-ME,300µMofeachdNTP,5mMactivatedDNA,andenzyme.

Unit definition:Oneunitistheamountofenzymerequiredtocata-lyzetheincorporationof10nmoloftotalnucleotideintoacidinsolubleformin30minutesat37°Cunderstandardassayconditions.


5X T7 DNA Polymerase Reaction Buffer (1 ml included, PN 70097):200mMTris-HCl(pH7.5),100mMMgCl2,and250mMNaCl.

T7 DNA Polymerase Dilution Buffer (1 ml included):20mMpotassiumphosphatebuffer(pH7.4),1mMDTT,0.1mMEDTA,50%glycerol.


References:1.Grippo,P.andRichardson,C.C.(1971)J. Biol.

Chem.246,6867-6873.2.Modrich,P.andRichardson,C.C.(1975)J. Biol.

Chem.250,5515-5522.3.Adler,S.andModrich,P.(1979)J. Biol. Chem.

254,11605-11614.4.Hori,K.,Mark,D.F.andRichardson,C.C.(1979)

J. Biol. Chem.254,11598-11604.5.Engler,M.J.,Lechner,R.L.andRichardson,C.C.

(1983)J.Biol.Chem.258,11165-11173.6.Lechner,R.L.andRichardson,C.C.(1983)

J. Biol. Chem.258,11185-11196.7.Bebenek,K.et al.(1989)Nucl. Acids Res. 17,5408.

T7 DNA Polymerase (E.C.2.7.7.7)



70052 200 units 1,000 units

Source:E. colistrainexpressingafulllengthunmodifiedcloneofTthDNAPolymerasefromThermus ther-mophilusstrainHB8.

Description:TthDNAPolymeraseisathermostableenzymederivedfromThermus thermophilus(1)whichhasbeenclonedandpurifiedfromE. coli.Theenzymepossessesahighlyprocessive5'→3'polymeraseactivitybutlacksa3'→5'exonucleaseactivity.TthDNAPolymeraseissuitableforPCRinthepres-enceofMg2+ions.ItalsohasanintrinsicreversetranscriptaseactivityinthepresenceofMn2+ions.ThereversetranscriptionactivityusingRNAasatemplatetopreparecDNAmaybeachievedathightemperatures,therebyeliminatingproblemsrelatedtosecondarystructure.TthDNAPolymeraseisverystablewithahalf-lifeof20minutesat95°C.

TthDNAPolymeraseissuitableforRT-PCRorforPCRalone.Theenzymeissuppliedwith5XRTBuffer,5XChelateBuffer,10XPCRBuffer,andseparatetubesof9mMMnCl2and25mMMgCl2.

Applications:�� Hightemperaturereversetranscriptionbetween60°-70°Ctohelpeliminatesecondarystructure�� QuantitationofviralRNAinclinicalsamples(2)

�� DetectionofmRNAinindividualcellsbyRT-PCR(3)

�� RapiddetectionofbacterialDNAsamplesbyPCRwithoutDNAextractionorcelllysis(4)

�� RT-PCRinthepresenceof2-5%(vol/vol)ofphenolsaturatedbuffer(5)

Properties:MolecularWeight:94kDaActivator:Mn2+forRNAdependentDNApolymerase

activity;Mg2+forDNAdependentDNApolymeraseactivity.

Purity:Greaterthan95%pureasdeterminedbySDS-PAGE.Freefromdetectablenon-specificendoribonucleases.

Storage buffer:20mMTris-HCl(pH8.5),1mMDTT,0.1mMEDTA,100mMKCI,50%glycerol,stabilizers.

Assay conditions:Thereactionmixturecontains67mMTris-HCl,pH8.8(at25°C),16.6mM(NH4)2SO4,6.7mMMgCl2,10mMβ-ME,200µMeachdATP,dGTP,dTTP,100µMdCTP,radio-labeleddCTP,250µg/mlactivat-edfishspermDNA,andTthDNAPolymerase.Afterincubationat74°Cfor10minutes,acidinsolublematerialisdetermined(50µlreactionvolume).

Unit definition:Oneunitincorporates10nmoloftotalnucleotidesintoacid-insolublematerialin30minutesat74°Cinatotalvolumeof50µl.


Functional test:TthDNAPolymeraseisfunctionallytestedforprod-uctyieldandlengthinRT-PCRandPCRamplifica-tion.

5X RT Buffer (included): 50mMTris-HCl,pH8.9,450mMKCl.

5X Chelate Buffer (included): 50mMTris-HCl,pH8.9,500mMKCl,3.75mMEGTA,0.25%Tween20,7.5mMMgCl2,25%glycerol.

10X PCR Reaction Buffer (included): 100mMTris-HCl,pH8.6,500mMKCl,15mMMgCl2.

25 mM MgCl2 (included): MgCl2 solution.

9 mM MnCl2 (included): MnCl2solution.


References:1.Ruttmann,C.M.,Corotas,M.,Zalvidar,J.and

Vicuna,R.(1985)Eur. J. Biochem.149,41-46.2.Mulder,J.,McKinney,N.,Christopherson,C.,

Sninsky,J.,Greenfield,L.andKwok,S.(1994)J. of Clinical Microbiology32,292-300.

3.Chiocchia,G.andSmith,K.A.(1997)BioTechniques 22,312-318.

4.Lofstrom,C.,Knutsson,R.,Axelsson,C.E.andRadstrom,P.(2004)Appl. and Environmental Microbiology.70,69-75.

5.Katcher,H.AndSchwartz,I.(1994)BioTechniques 16,84-92.

Tth DNA Polymerase

Fig. 1. RT/PCR of target with high G+C content (77%) 0.387 kb Notch3 (lane1) and 1.0 kb single copy gene Numb (lane 2).

Fig. 2. PCR of DNA fragments using Tth DNA Polymerase ranging from 1 kb to 6 kb from Lambda DNA.

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Molecular Biology Enzymes | Polymerases—Reverse Transcriptases


Source:PurifiedvirionofAvian myeloblastosisvirus.

Description:AMVReverseTranscriptasecatalyzesthepolymeriza-tionofDNAusingtemplateDNA,RNAorRNA:DNAhybrids(1).Itconsistsoftwopolypeptidechains,oneofwhichcontainsa5'→3'polymeraseactivityandtheotheranRNaseHactivity.ReversetranscriptaserequiresMg2+orMn2+andisusedforfirststrandsynthesisofcomplementaryDNA(cDNA)frommRNAtemplate(2).Underproperconditions,highyieldsoffull-lengthcDNAcanbeobtainedwithAMVReverseTranscriptase(3).AMVReverseTranscriptaseisalsousefulintheamplificationofRNA(4).

Applications:�� SynthesisofcDNAforPCR,cloning,andhybridizationprobes�� Filling-inandlabelingthe3'terminiofDNAwith5'protrudingends�� RNAsequencing�� AmplificationofRNA(4)

�� Primerextensionassays

Properties:MolecularWeight:157kDaInactivation:75°Cfor10minutesorbyadding2µl

of0.5MEDTAfora50µlreaction.

Purity:Testedforcontaminatingendonucleases,exonucle-asesandribonucleases.

Storage buffer:200mMpotassiumphosphate(pH7.2),2mMDTT,0.2%TritonX-100,50%glycerol.

Assay conditions:Thereactionmixture(100µl)contains50mMTris-HCI(pH8.3),6mMMgCl2,40mMKCI,0.5mMradiolabeledTTP,and0.4mMPoly(rA)n:oligo(dT)50.Incubationisat37°Cfor10minutes.

Unit definition:Oneunitistheamountofenzymerequiredtoincor-porate1nmolofradiolabelednucleotideintoacidinsolubleproductin10minutesat37°C.


Functional test:RT-PCRamplificationongenerationof0.5kband1.5kbRT-PCRproducts.

Functionally tested 5X AMV RT Reaction Buffer (1 ml included):250mMTris-HCl(pH8.3),40mMMgCl2,250mMNaCl,5mMDTT

Shipping and storage:Shippedondryice.Storeat-20°C.Avoidfreeze-thawcycles,whichresultinlossofenzymeactivity.

References:1.Kacian,D.L.(1977)inMethods in Virology,eds.

K.MaramoroschandH.Koprowski6,143-184.2.Goodman,H.M.andMacDonald,R.J.(1979)

Methods Enzymol.68,75-90.3.Berger,S.L.,Wallace,D.M.,Puskas,R.S.and

Eschenfeldt,W.H.(1983)Biochemistry22,2365-2372.

4.PCRProtocols:AGuidetoMethodsandApplications(1990)eds.M.A.Innis,D.H.Gelfand,andJohnJ.Sni,(AcademicPress,NewYork),23.

AMV Reverse Transcriptase

78306 25,000 units 100,000 unitsCustoms by request

M-MLV Reaction Buffer, 5X(Included with each pack size of enzyme)71505 1 ml

Source:E. colistraincontaininganoverproducingcloneofM-MLVReverseTranscriptase.

Description:M-MLVReverseTranscriptasecatalyzesthepolymerizationofDNAusingtemplateDNA,RNA,orRNA:DNAhybrids(1).Full-lengthcopiesoflargemRNAs,>10kb,maybesynthesized.M-MLVReverseTranscriptasehasamuchlowerRNaseHactivitythanAMVReverseTranscriptase,resultinginhighyieldsoffulllengthcDNA(2).ThismakesM-MLVReverseTranscriptaseveryusefulincDNAsynthesisandRT-PCR.

ForaddedconvenienceandoptimalRT-PCR,trytheRT-PCRKits(PN78350and78355)andRT-PCRMasterMixes(PN71185and78370).

Applications:�� RT-PCR�� SynthesisoffirststrandcDNAforPCR,cloning,andhybridizationprobes�� Filling-inandlabelingthe3'terminiofDNAwith5'protrudingends�� AmplificationofRNA�� Primerextensionassays

Properties:MolecularWeight:71kDa(monomeric).Inhibitors:Polyamines,phosphate,pyrophosphates,

andlithiumchloride(3,4).Inactivation:75°Cfor10minutesorbyadding2µl

of0.5MEDTAfora50µlreaction.

Purity:Greaterthan90%pureasdeterminedbySDS-PAGE.Testedforcontaminatingendonucleases,exonucle-ases,andribonucleases.

Storage buffer:20mMTris-HCl(pH7.5),0.1MNaCl,0.1mMEDTA,1mMDTT,0.01%IgepalCA-630,50%glycerol.

Assay conditions:Thereactionmixture(25µl)contains50mMTris-HCI(pH8.3),40mMKCl,6mMMgCl2,1mMDTT,400µMpoly(rA)-oligo(dT)12-18,500µMradiolabeledTTP,and0.1-0.5unitsenzyme.Incubationisat37°Cfor10minutes.

Unit definition:Oneunitisthatamountofenzymerequiredtoincorporate1nmolofdeoxynucleotideintoDE-81filter-bindingmaterialin10minutesat37°Cusingpoly(rA)-oligo(dT)12-18astemplate-primer.


Functional test:RT-PCRamplificationongenerationof0.5kband1.5kbRT-PCRproducts.

Functionally tested 5X M-MLV Reaction Buffer (1 ml included, PN 71505):250mMTris-HCl(pH8.3),375mMKCl,15mMMgCl2,50mMDTT.


References:1.Roth,M.J.,Tanese,N.andGoff,S.P.(1985)

J.Biol.Chem.260,9326-9335.2.Sambrook,J.andRussell,D.W.(2001)Molecular

Cloning: A Laboratory Manual,3rdEdition,ColdSpringHarbor,NewYork,8.48.

3.Gerard,G.F.andD’Alessio,J.M.(1993)Methods In Molecular Biology16, HumanaPress,NJ,73-93.

4.Sambrook,J.andRussell,D.W.(2001)Molecular Cloning: A Laboratory Manual,3rdEdition,ColdSpringHarbor,NewYork,A8.16.

M-MLV Reverse Transcriptase(E.C.2.7.7.49)


Molecular Biology Enzymes | Polymerases—RNA Polymerases


Source:E. coliRNAPolymeraseHoloenzymeisisolatedfromtherifampicin-sensitivestrainBL21andissigmasaturated.

Description:E. coliRNAPolymeraseisaholoenzymeusedfortranscriptionofDNAandtheproductionoflabeledRNAforhybridizationprobes.Thepolymerasecon-sistsof5subunitsdesignateda,a,β',β,andσ.Theenzymeis100%saturatedwithsigmafactorwhichisrequiredforcorrectinitiationofRNAsynthesisonDNAtemplatescontainingbacterialandphagepromoters.

Application:�� SynthesisoftranscriptswhenitisnotpossibletotranscribeDNAfromavectorcontainingaphageRNApromoter

Properties:MolecularWeight:Approximately450kDa.Themolecularweightforthesubunitsare36.5(each),156,151,and70kDafora,β',β,andσsubunitsrespectively.

Purity:Testedforcontaminatingnon-specificexonucleases,endonucleases,andribonucleases.

Storage buffer:50mMTris-HCl(pH7.5),250mMNaCl,0.1mMEDTA,1.0mMDTT,50%glycerol.

Assay conditions:Thereactionmixture(250µl)contains40mMTris-HCI(pH7.5),150mMKCI,10mMMgCl2,0.01%TritonX-100,10mMDTT,0.02µg/µlDNAtemplate,0.5mMofeachrNTP,andenzyme.Incubationisat37°Cfor10minutes.

Unit definition:Oneunitistheamountofenzymerequiredtocata-lyzetheincorporationof1nmolofaribonucleosidetriphosphateintoRNAin10minutesat37°C.

Concentration:1unit/µl


References:1.Burgess,R.R.andTravers,A.A.(1970)Fed Proc.

29,1164-1169.2.Burgess,R.R.,Travers,A.A.,Dunn,J.J.andBautz,

E.K.(1969)Nature221,43-46.3.Fujiki,H.,Palm,P.,Zillig,W.,Calendar,R.and

Sunshine,M.(1976)Mol.Gen.Genet.145,19-22.

E. coli RNA Polymerase Holoenzyme(RNANucleotidylTransferase,E.C.2.7.7.6)


Poly(A) Polymerase Reaction Buffer, 5X(Included with each pack size of enzyme)74226 1 ml

Source:E. colistraincontaininganoverproducingcloneofYeastPoly(A)Polymerase.

Description:Poly(A)Polymerasecatalyzestheadditionofadeno-sineresiduesontothe3'-endsofRNA(1,2).Itcanbeusedtoaddpoly(A)tailstoRNAinthefirststepofcloning(3).ThereactionrequiresMn2+orMg2+,ATPassubstrate,andanyRNAcontaining3'hydroxylter-miniasprimer.LongerRNAmoleculesaresomewhatbetterprimersthanshortoligomers(4).Substitutionofcordycepin-5'-triphosphate(3'-dATP)forATPresultsinadditionofasingle3'-dAresiduetotheendsoftheRNA,ausefultechniqueforlabelingRNAatthe3'-end(5)*.

Incomparativestudies,YeastPoly(A)PolymeraseworksmoreefficientlythanE. coliPoly(A)Polym-eraseforRNAoligonucleotide-labelingandpoly(A)tailing.Shorterincubationtimesarerequiredfortheyeastenzymeanditisfoundtolabelbothlongandshortsubstratesequallywell.Poly(A)PolymeraseisrecommendedoverT4RNALigasefor3'-endlabel-ingoflongRNAmolecules.

*Note:Variousmodifiednucleotidescanalsobeusedtolabelthe3'endofRNAusingYeastPoly(A)Polymerase(5).

Applications:�� Additionofpoly(A)tailstoRNA�� Labelingthe3'endsofRNA

Purity:Ribonucleasefree.

Storage buffer:20mMTris-HCl(pH8.0),50mMKCl,0.5mMDTT,50%glycerol.

Assay conditions:25mMTris-HCl(pH7.0),40mMKCl,0.5mMMnCl2,0.5mMEDTA,0.5mMDTT,0.2mg/mlBSA,10%glycerol,3.3µMradiolabeledATP,0.5mMATP,6.5µgpoly(A)(~100bases),poly(A)polymerase.Afterincubationat37°Cfor10minutes,acidinsolubleradioactivityisdetermined.

Unit definition:Oneunitistheamountofenzymewhichincorpo-rates1pmolAMPintoacid-insolublematerialat37°Cin1minute.


Functional test:3'-endlabelingofaribonucleotidewithcordycepin-5'-triphosphate.

Functionally tested 5X Poly(A) Polymerase Reaction Buffer (1 ml included, PN 74226):100mMTris-HCl(pH7.0),3.0mMMnCl2,0.1mMEDTA,1mMDTT,500µg/mlacetylatedBSA,50%glycerol.


References:1.Sippel,A.E.(1973)Eur.J.Biochem.37,31-40.2.Edmonds,M.(1982)inThe Enzymes,3rdedition,

ed.P.D.Boyer(AcademicPress,NewYork)15,217-244.

3.Gething,M.J.,Bye,J.,Skehel,J.andWaterfield,M.(1980)Nature 287,301-306.

4.Sano,H.andFeix,G.(1976)Eur.J.Biochem.71,577-583.

5.Martin,G.andKeller,W.(1998)RNA4,226-230.

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Molecular Biology Enzymes | Polymerases—RNA Polymerases

Standard concentration, 20 units/µl70047Y 6,000 unitsHigh concentration, >80 units/µl70001Z 30,000 unitsCustoms by request

Source:E. colistraincontaininganoverproducingcloneofT7RNAPolymerase(1).

Description:T7RNAPolymeraseisasingle-subunitenzymeproducedbybacteriophageT7(2).ItishighlyspecificforT7promoterandterminatorsequences(2,3).Ithasbeenwidelyusedfortherapidsynthesisin vitroofspecificRNAs.ThesetranscriptscanbeuseddirectlyassubstratesforstudiesofRNAstructureorme-tabolism.Ifthetranscriptsaresuitablylabeled,theycanalsobeusedassensitivehybridizationprobes.T7RNAPolymerase,alongwithchain-terminatingnucleosidetriphosphates(4),havealsobeenusedforthedirectsequencingofDNA.T7RNAPolymeraseisalsousedtogeneratecappedmRNAforexpressionstudiesinoocytesandothercells(5).

Applications:�� ProductionofRNAtranscriptsforhybridizationprobes�� ProductionoflargeamountsofdiscretesizedRNA�� ProductionofcappedormodifiedRNAtranscripts

Properties:MolecularWeight:98.8kDaOptimumpH:7.7-8.3OptimumTemperature:37°CRequirementforDivalentCation:Mg2+(optimal

concentrationis20mM)MichaelisConstants(2):40µMATP,160µMGTP,

60µMUTP,80µMCTP

Purity:Greaterthan99%pureasdeterminedbySDS-PAGE.Testedforcontaminatingnonspecificendonucleases,exonucleases,andribonucleases.

Storage buffer:20mMHPO4,pH7.7,100mMNaCl,1mMEDTA,10mMDTT,1%CHAPS,and50%glycerol.

Assay conditions:Thereactionmixturecontains1XTranscriptionBuffer,0.5mMeachNTP,1µCilabeledCTP,1µgofT7promotercontainingDNA,andT7RNAPolymerase.Incubationisat37°Cfor15minutes(50µlreactionvolume).

Unit definition:Oneunitcatalyzestheincorporationof1nmolofCMPintopolynucleotidefractionboundtoapositivelychargedDE-81filterpaperin60minutesat37°CusingalinearizedT7promotercontaining280nttemplate.

Concentration:Standardconcentration(PN70047Y):20units/µlHighconcentration(PN70001Y/Z):>80units/µl

Functional test:TranscriptionfromplasmidcontainingT7promoter;>10µgofRNAcanbeproducedfrom1µgofsupercoiledtemplate.

Functionally tested 10X T7/T3 RNA Polym-erase Transcription Buffer (included):400mMTris-HCl,pH8.0,60mMMgCl2,100mMDTT,100mMNaCl,20mMspermidinetrihydro-chloride.


References:1.Tabor,S.andRichardson,C.C.(1985)Proc.Natl.

Acad.Sci.USA 82,1074-1078.2.Chamberlin,M.andRingJ.(1973)J.Biol.Chem.

248,2235-2244,2245-2250.3.Chamberlin,M.andRyan,T.(1982)inThe

Enzymes,3rdedition,ed.P.D.Boyer(AcademicPress,NewYork.) 15,87-108.

4.Axelrod,V.D.andKramer,F.R.(1985)Biochemistry 24,5716-5723.

5.Sambrook,J.andRussell,D.W.(2001)Molecular Cloning: A Laboratory Manual,3rdEdition,ColdSpringHarbor,NewYork,9.87-9.88.

6.Ausubel,F.M.,Brent,R.,Kingston,R.E.,Moore,D.D.,Seidman,J.G.,Smith,J.A.andStruhl,K.(1998)Current Protocols in Molecular Biology(JohnWileyandSons,Inc.).

T7 RNA Polymerase (RNANucleotidylTransferase,E.C.2.7.7.6)

Standard concentration, 20 units/µl70051Y 1,000 units70051Z 5,000 unitsHigh concentration, 200 units/µl70204 1,000 units 5,000 units

Source:E. coli straincontaininganoverproducingcloneofT3RNAPolymerase.

Description:T3RNAPolymerasehasahighspecificityforbacteriophageT3promotersequences.ItisusedinthesynthesisoflargeamountsofspecificRNAtranscriptsfromvectorscontainingtheT3promoter.TranscriptsmaybeusedasprobesinNorthernandSouthernblots(1),substratesforin vitro translationstudies(2),substratesforRNAprocessingstudies(3),andforexonandintronmappingofgenomicDNA(1).Theenzymemaybeusedwithbothradioactivelylabeled(4)andhapten-labeled(fluorescein,digoxigen-in,biotin,etc.)nucleosidetriphosphates(5).T3RNAPolymeraseisalsousedtogeneratecappedmRNAforexpressionstudiesinoocytesandothercells(6).

Applications:�� ProductionofRNAtranscripts�� ProductionofcappedormodifiedRNAtranscripts


Storage buffer:20mMHPO4(pH7.7),100mMNaCl,1mMEDTA,10mMDTT,1.0%CHAPS,50%glycerol.

Assay conditions:Thereactionmixturecontains1XTranscriptionBuffer,0.5mMeachNTP,1µCilabeledCTP,1µgofT3promotercontainingDNA,andT3RNAPolymerase.Incubationisat37°Cfor15minutes(50µlreactionvolume).

Unit definition:Oneunitcatalyzestheincorporationof1nmolofCMPintopolynucleotidefractionboundtoapositivelychargedDE-81filterpaperin60minutesat37°CusingalinearizedT7promotercontaining280nttemplate.

Concentration:Standardconcentration(PN70051Y/Z)20units/µlHighconcentration(PN70204)200units/µl

Functionally tested 10X T7/T3 RNA Polymerase Transcription Buffer (included):400mMTris-HCl(pH8.0),60mMMgCl2,100mMDTT,100mMNaCl,20mMspermidinetrihydro-chloride.


References:1.Melton,D.A.,Krieg,P.A.,Regagliati,M.R.,

Maniatis,T.,Zinn,K.andGreen,M.R.(1984)Nucl.Acids Res.12,7035-7056.

2.Krieg,P.A.andMelton,D.A.(1984)Nucl.Acids Res.12,7057-7070.

3.Green,M.R.,Maniatis,T.andMelton,D.A.(1983)Cell 32,681-694.

4.Davanloo,P.,Rosenberg,A.H.,Dunn,J.J.andStudier,F.W.(1984)Proc.Natl.Acad.Sci.USA 81,2035-2039.

5.Langer,P.R.,Waldrop,A.A.andWard,D.C.(1981)Proc.Natl.Acad.Sci.USA 78, 6633-6637.

6.Sambrook,J.andRussell,D.W.(2001)Molecular Cloning: A Laboratory Manual,3rdEdition,ColdSpringHarbor,NewYork,9.87-9.88.

T3 RNA Polymerase (RNANucleotidylTransferase,E.C.2.7.7.6)


Molecular Biology Enzymes | Polymerases—Transferase

72033 500 units 2,500 unitsCustoms by request

Source:E. colicloneexpressingbovineTdT

Description:TerminalDeoxynucleotidylTransferase(TdT)catalyzesthetemplate-independentadditionofunlabelledorradio-,fluorescent-,biotin-,ordigoxygenin-labelleddeoxynucleotides,dideoxynucleotides,orribonucleo-tidestothe3'-hydroxylterminiofsingle-ordouble-strandedDNA.TdTrequiresanoligodeoxy-nucleotideprimerwithatleastthreephosphategroupsandafree3'-hydroxylendandadivalentcationforactivity.TdTalsocatalyzesthedepolymer-izationofDNA(pyrophosphorolysis)andpyrophos-phateexchange(1,2).

Historically,TdTisusedtolabelDNAprobesinapop-tosisdetectionandquantificationintheTdTdUTPnickendlabelingmethod(TUNEL)(alsoreferredtoasISEL,in situendlabeling),inDNAsequencing,(1,3)andtocreate“sticky”endsby3'tailingDNApriortoannealingandtransformation(4).For5'RACE,TdTisusedtoaddhomopolymertailstothe3'-endofthefirststrandcDNA(5).Recently,TdThasbeenusedtocloneunknownsequencesadjacenttoknownsequencesingenomicDNA(6)andforTdT-dependentPCR(7,8)(TDPCR).TDPCRisusedinRNAanaly-sis(9,10,11),transcriptionanalysis(12,13),DNAdamageanalysis(14),andDNAfootprinting(7).

Applications:�� 5'RACE�� Creatingisotopic,fluorescent,biotinylated,ordigoxygeninlabeledprobes�� Creatingfluorescent3'labeledsequencingprimers�� EliminationofbackgroundbandsinDNAsequencing�� TDPCR�� DetectionofapoptoticDNAdamage(TUNELassay)


Purity:Testedforcontaminatingexonucleasesandendo-nucleases.

Storage buffer:50mMpotassiumphosphate,pH6.4,100mMNaCl,1mMDTT,0.1%Tween20,50%glycerol.

Assay conditions:Thereactionmixture(50µl)containing25mMsodiumcacodylate(pH7.2),0.5mMradiolabelleddATP,10mMMgCl2,0.1mMβ-ME,0.01mMd(pA)35,andenzymeisincubatedat37°Cforvaryingamountsoftime.

Unit definition:Oneunitistheamountofenzymerequiredtopolymerize1nmolofdATPresiduesintoDNAin60minutesat37°Cunderstandardassayconditions.


Functional test:Functionallytestedby3'-endlabelingofafluores-centlylabeledoligonucleotide.

Functionally tested 5X TdT Reaction Buffer (1 ml included):500mMsodiumcacodylate,pH6.8,5mMcobaltchloride,0.5mMDTT.


References:1. Anderson,R.S.,Bollum,F.J.,andBeattie,K.L.

(1999)Nucl. Acids Res.27,3190-3196.2. Krayevsky,A.A.,Victorova,L.S.,Arzumanov,A.

A.,andJasko,M.V.(2000)Pharm & Therapeutics85,165-173.

3. TechTip201,UseofTerminalDeoxynucleotidylTransferase(TdT)toresolveBAFLs,USBCorporation.

4. Sambrook,J.andRussell,D.W.(2001)“MolecularCloning:ALaboratoryManual,”ColdSpringHarborLaboratoryPress,ColdSpringHarbor,NY,11.110.

5. Sambrook,J.andRusselL,D.W.(2001)“MolecularCloning:ALaboratoryManual,”ColdSpringHarborLaboratoryPress,ColdSpringHarbor,NY,8.54-8.60.

6. Liu,X.andBaird,W.V.(2001)Plant Molec. Biol. Reporter19,261-267.

7. Komura,J.andRiggs,A.D.(1998) Nucl. Acids Res.26,1807-1811.

8. Davies,N.P.andMurray,V.(2000)BioTechniques29,1168-1172.

9. Schmidt,W.M.andMueller,M.W.(1996)Nucl. Acids Res.24,1789-1791.

10.Albuquerque-Silva,J.,Houard,S.,andBollen,A.(1998)Nucl. Acids Res.26,3314-3316.

11.Chen,H.,Castanotto,D.,Lebon,J.M.,Rossi,J.J.andRiggs,A.D.(2000)Nucl. Acids Res.28,1656-1664.

12.Buettner,V.L.,Lebon,J.,Gao,C.,Riggs,A.D.,andSinger-Sam,J.(2000)Nucl. Acids Res.28,e25.

13.Tagoh,H.,Himes,R.,Clarke,D.,Leenen,P.,Riggs,A.D.,Hume,D.,andBonifer,C.(2002)Genes & Development16,1721-1737.

14.Arlt,V.M.,Schmeiser,H.H.,andPfeifer,G.P.(2001)Carcinogenesis22,133-140.

Related products – Ribonucleotides


Guanosine-5'-Triphosphate (GTP), 100 mM Solution77243 25 µmol

Related products – Deoxynucleotides

2'-Deoxyadenosine-5'-Triphosphate (dATP) Sodium Salt, 100 mM Solution77102 25 µmol

2'-Deoxycytidine-5'-Triphosphate (dCTP) Sodium Salt, 100 mM Solution77104 25 µmol

2'-Deoxyguanosine-5'-Triphosphate (dGTP) Sodium Salt, 100 mM Solution77106 25 µmol

2'-Deoxythymidine-5'-Triphosphate (dTTP) Sodium Salt, 100 mM Solution77108 25 µmol

Related products – Dideoxynucleotides

2',3'-Dideoxyadenosine-5'-Triphosphate (ddATP), 10 mM Sodium Salt Solution77110 1 µmol

2',3'-Dideoxycytidine-5'-Triphosphate (ddCTP), 10 mM Sodium Salt Solution77112 1 µmol

2',3'-Dideoxythymidine-5'-Triphosphate (ddTTP), 10 mM Sodium Salt Solution77116 1 µmol

Terminal Deoxynucleotidyl Transferase, Recombinant (rTdT)(E.C.2.7.7.31)


Molecular Biology Enzymes | RNase Inhibitors


Source:Humanplacenta

Description:HumanPlacentalRNaseInhibitor(HPRI)bindsandinhibitsabroadrangeofeukaryoticRNases,includ-ingRNaseA,RNaseB,RNaseC,andhumanplacen-talRNase.HPRIispurifiedfromhumanplacentabyaffinitychromatographyonanimmobilizedRNaseAcolumn.Itformsa1:1complexwithRNaseAandinhibitsRNaseactivitynon-competitively(Ki=3x1010M).ItcanbeaddeddirectlytoreactionmixturescontainingRNA.HPRIdiffersfromothercompetitiveinhibitorsinthatitcanbeeasilyremovedfromreac-tionmixturesbyphenolextraction.

RNaseInhibitorisnoteffectiveagainstRNase1,RNaseT1,S1Nuclease,RNaseH,orRNasefromAspergillus.Tomaintainactivity,HPRIrequiresaminimumof1mMDTTandisactiveoverabroadpHrange(pH5.5-9.0).

Applications:�� cDNAsynthesis(2)

�� In vitrotranslation(3)

�� In vitrotranscription(4)

�� Polysomeisolation�� RT-PCR�� IsolationofmammaliancellsamplesthatcontainamRNA-proteincomplex(3)

�� Separationandidentificationofspecificribonucleaseactivities(5)

Properties:Activators:RequiresDTTforactivityInhibitors:RNaseswillrecoveractivityafter

inactivationattemperaturesgreaterthan55°Corin7Murea.

Purity:Testedforcontaminatingnon-specificendonu-cleases,exonucleases,ribonucleases,andlatentribonucleases.

Storage buffer:20mMHEPES-KOH(pH7.6),50mMKCl,8mMDTT,50%glycerol.

Unit definition:Oneunitinhibitstheactivityof5ngofRNaseAby50%(1).Thisinhibitoractivityisdeterminedbyitsabilitytoinhibitthehydrolysisofcyclic2'3'-CMPbyRNaseA.

Concentration:≥20units/µl

Shipping and storage:Shippedondryice.Storeat-20°C.Avoidrepeatedfreeze/thawcycles.

References:1.Blackburn,P.(1979)J. Biol. Chem.254,12484-

12487.2.deMartynoff,G.,Pays,E.andVassart,G.(1980)

Biochem. Biophys. Res. Commun.93,645-653.3.Scheele,G.andBlackburn,P.(1979)Proc. Natl.

Acad. Sci. USA76,1898-1902.4.Nielson,D.A.andShapiro,D.J.(1986)Nucl. Acids

Res.14,5936.5.Eichler,D.C.,Tatar,T.F.andLasater,L.S.(1981)

Biochem. Biophys. Res. Commun.101,396-403.

RNase Inhibitor (Human Placenta)


Source:E. colistraincontaininganoverproducingcloneofhumanplacentaribonucleaseinhibitor.

Description:RecombinantHumanPlacentalRNaseInhibitor(rHPRI)bindsandinhibitsabroadrangeofeukary-oticRNases,includingRNaseA,RNaseB,RNaseCandhumanplacentalRNase.HPRIformsa1:1complexwithRNaseAandinhibitsRNaseactivitynon-competitively(Ki=3x1010M).ItcanbeaddeddirectlytoreactionmixturescontainingRNA.HPRIdiffersfromothercompetitiveinhibitorsinthatitcanbeeasilyremovedfromreactionmixturesbyphenolextraction.

RNaseInhibitorisnoteffectiveagainstRNase1,RNaseT1,S1Nuclease,RNaseH,orRNasefromAspergillus.Tomaintainactivity,HPRIrequiresaminimumof1mMDTTandisactiveoverabroadpHrange(pH5.5-9.0).

Applications:�� cDNAsynthesis(2)

�� In vitrotranslation(3)

�� In vitrotranscription(4)

�� Polysomeisolation�� RT-PCR�� IsolationofmammaliancellsamplesthatcontainamRNA-proteincomplex(3)

�� Separationandidentificationofspecificribonucleaseactivities(5)

Properties:MolecularWeight:50kDaIsoelectricPoint:4.7Activators:DTTInhibitors:RNaseswillrecoveractivityafter

inactivationattemperaturesgreaterthan55°Corin7Murea.


Storage buffer:20mMHEPES-KOH(pH7.6),50mMKCl,8mMDTT,50%glycerol.

Unit definition:Oneunitinhibitstheactivityof5ngofRNaseAby50%(1).


Shipping and storage:Shippedondryice.Storeat-20°C.Avoidrepeatedfreeze/thawcycles.

References:1.Blackburn,P.(1979)J. Biol. Chem.254,12484-

12487.2.deMartynoff,G.,Pays,E.andVassart,G.(1980)

Biochem. Biophys. Res. Commun.93,645-653.3.Scheele,G.andBlackburn,P.(1979)Proc. Natl.

Acad. Sci. USA76,1898-1902.4.Nielson,D.A.andShapiro,D.J.(1986)Nucl. Acids

Res.14,5936.5.Eichler,D.C.,Tatar,T.F.andLasater,L.S.(1981)

Biochem. Biophys. Res. Commun.101,396-403.

RNase Inhibitor (Recombinant)


Molecular Biology Enzymes | Topoisomerase

78303Y 200 units 78303Z 500 unitsCustoms by request

Source: E. coli containingacloneofthehumanTopoisomer-aseIIgene.

Description:TopoisomeraseIIaltersthetopologicalstateofnucleicacidsbypassinganintactDNAhelixthroughatransientbreakwhichgeneratesaseparateDNAhelix(1,2).Asaresultofitsdouble-strandedDNApas-sagemechanism,theenzymecanrelaxnegativelyorpositivelysupercoiledDNA,aswellascatenate/decatenateorknot/unknotDNAmolecules.Topoi-someraseIIhasanabsoluterequirementfordivalentcationandATP(ordATP).

Applications:�� RelaxnegativelyorpositivelysupercoiledDNA�� CatenatingordecatenatingDNA�� KnottingorunknottingDNA

Properties:MolecularWeight:340kDahomodimerOptimumTemperature:30°-37°COptimumpH:7.9RequirementsforDivalentCation:5mMMg2+

forcatalyticactivityand5mMCa2+forDNAcleavage.

OptimumATPConcentration:0.5to1.0mMOptimumIonStrength:100-170mMKCl/NaClInhibitors:N-ethylmaleimide,zinc,novobicin,

coumermycinA1,etoposide,amsacrine

Purity:Greaterthan98%pureasdeterminedbysilver-stainedSDS-PAGE.Freeofcontaminatingexonucle-asesandendonucleases.

Storage buffer:15mMsodiumphosphate(pH7.1),700mMNaCl,0.1mMEDTA,0.5mMDTT,50%glycerol.

Assay conditions: Thereactionmixturecontains10mMTris-HCl,pH7.9,175mMKCl,0.1mMEDTA,5mMMgCl2,2.5%glycerol,1mMATP.

Unit definition:Oneunitistheamountofenzymerequiredtofullyrelax0.3µg(5nM)ofnegativelysupercoiledpBR322plasmidDNAin15minutesat30°Cunderthestandardassayconditions.


Functional test:TestedforactivityinastandardTopoisomeraseIIunitassay.

Functionally tested 10X Topoisomerse II Reaction Buffer (1 ml included):100mMTris-HCl(pH7.9),500mMNaCl,500mMKCl,50mMMgCl2,1mMEDTA,150µg/mlBSA,10mMATP.

Topoisomerase II Dilution Buffer (1 ml included):10mMsodiumphosphate(pH7.1),50mMNaCl,0.2mMDTT,0.1mMEDTA,0.5mg/mlBSA,10%glycerol.

Shipping and storage:Shippedondryice.Storeat-20°Cforfrequentuse.Storeat-80°Cforlong-termstorage.

References:1.Bjergbaek,L.,Kingma,P.,Nielsen,I.S.,Wang,Y.,

Westergaard,O.,Osheroff,N.andAnderson,A.H.(2000)J. Biol. Chem.275,13041-13048.

2.Bromberg,K.D.,Hendricks,C.,Burgin,A.B.andOsheroff,N.(2002)J. Biol. Chem.277,31201-31206.

3.Renodon-Corniere,A.,Jensen,L.H.,Nitiss,J.L.,Jensen,P.B.andSehested,M.(2002)Biochemistry41,13395-13402.

4.Sabourin,M.andOsheroff,N.(2000)Nucl. Acids. Res.28,1947-1954.

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Topoisomerase II, Alpha (DNARelaxingEnzyme)

Molecular Biology Products & KitsGene Regulation

Transfection

Nuclear Extraction

PCR Methods

Transcription Factors

Stable Cell Lines

Cellular Manipulation

NGS Library Prep

Cloning

Mutagenesis

Labeling

DNA

Markers

Protein Analysis

Proteinases


Molecular Biology Products & Kits | Table of Contents

Gene Regulation - TransfectionDeliverX™ and DeliverX Plus siRNA

Transfection Kits 98

Cellular ManipulationDeliverX Peptide Transfection Kits 99Cancer Cell Isolation Kit 101

Gene Regulation – Nuclear ExtractionNuclear Extraction Kit 102

Gene Regulation – PCR MethodsVeriQuest™ qPCR and qRT-PCR Master Mixes 103HotStart-IT™ qPCR and qRT-PCR Master Mixes 103Promoter Methylation PCR Kit 103Poly(A) Tail-Length Assay Kit 103

Gene Regulation – Transcription FactorsEMSA (Gel-Shift) Kits and Probe Sets 103TF Activation ELISA Kits 107Gene Promoter Reporter Vectors 111Luciferase Reporter Vectors 112Mammalian Full-length TF Expression Vectors 114Function-Specific Protein/DNA Arrays 115TF-TF Interaction Arrays 115Protein/DNA Arrays 116NFkB-TF and PPAR-TF Interaction Arrays 118

Gene Regulation – Stable Cell LinesReporter Stable Cell Lines 119

Next Generation Sequencing (NGS) Library PrepPrep2Seq™ DNA Library Prep Kit for

Illumina® platform 123Prep2Seq Multiplex Oligo Adapters for

Illumina® platform 124

CloningLigate-IT™ Rapid Ligation Kit 125

MutagenesisChange-IT™ Multiple Mutation Site Directed

Mutagenesis Kit 126

LabelingSequenase™ Random Primer Labeling Kit 127

DNA 128

DNA MarkersPCR Markers, 50-2000 bp 130DNA Ladder, 100 bp 130DNA Ladder, 1 kb Plus 131

Oligonucleotide MarkerLow Molecular Weight Marker, 10 – 100 nt 132

Protein AnalysisAngiogenesis Antibody Arrays 133Cancer Antigen ELISA Kits 134Cytokine Antibody Arrays 134

Protein MarkersProtein Markers, 10-225 kDa 136

ProteinasesProteinase K 137Proteinase K Solution 137


Molecular Biology Products & Kits | Gene Regulation—Transfection

DeliverX siRNA Transfection Evaluation KitDX0001 0.12 mlDeliverX siRNA Transfection KitDX0002 0.4 mlDeliverX siRNA Transfection KitDX0003 1.0 mlDeliverX siRNA Transfection KitDX0004 4 x 1.0 mlDeliverX Plus siRNA Transfection Evaluation KitDX0051 0.12 mlDeliverX Plus siRNA Transfection KitDX0052 0.4 mlDeliverX Plus siRNA Transfection KitDX0053 1.0 mlDeliverX Plus siRNA Transfection KitDX0054 4 x 1.0 mlFAM-labeled siRNA ControlDX0100 0.12 mlHuman GAPDH siRNA ControlDX0101 0.06 mlDeliverX siRNA Buffer-1DX0500 4 mlDeliverX siRNA Buffer-1DX0501 14 mlDeliverX siRNA Buffer-1DX0502 34 mlDeliverX siRNA Buffer-2DX0503 4 mlDeliverX siRNA Buffer-2DX0504 14 mlDeliverX siRNA Buffer-2DX0505 34 mlDeliverX Plus siRNA Buffer-1DX0551 4 mlDeliverX Plus siRNA Buffer-1DX0552 14 mlDeliverX Plus siRNA Buffer-1DX0553 34 mlDeliverX Plus siRNA Buffer-2DX0554 4 mlDeliverX Plus siRNA Buffer-2DX0555 14 mlDeliverX Plus siRNA Buffer-2DX0556 34 ml

�� Novel peptide-based, non-endosomal delivery mechanism�� Efficient delivery of siRNA into difficult-to-transfect cell types�� High cell viability at optimal delivery conditions

DeliverX Transfection Reagents are based on novel “MPG” delivery technology developed at Centre de Recherches en Biochimie Macromoléculaire (CNRS) in Monpellier, France, in the laboratory of Dr. F. Heitz and Dr. G. Divita. MPG technology uses virus-derived amphipathic peptides that directly interact with nucleic acid cargos to form nanoparticles (150- 200 nm) capable of crossing through plasma

membranes and releasing their contents inside the cell. The mechanism of entry is receptor-independent, involves MPG/lipid interactions, and avoids the endocytic pathway, thereby preventing endosomal or lysosomal degradation of cargo. MPG peptides can be designed to accommodate specific molecular cargos including siRNAs, single and double-strand oligonucleotides, small peptides, morpholino antisense oligonucleotides, plasmids, and proteins.

DeliverX siRNA Reagents are available in two formats. DeliverX siRNA is a non-validated solution which comes with detailed instructions for carrying out assay optimization and can be used per any cell line of choice.

DeliverX Plus siRNA is a validated solution for spe-cific cell lines. With an initial offering of 18 validated cell lines, DeliverX Plus siRNA is a complete answer for your transfection needs, coming with a detailed protocol and certificate of analysis for each lot. It de-livers an assurance of success each and every time.

Regardless of which solution suits your needs, each of these reagents offers unrivalled efficiency, minimal toxicity, and a simple workflow.

Transfection proficiency controls Transfection proficiency controls are tools to enable users to optimize transfections under controlled conditions. These controls are available separately or as part of our evaluation kits.

Our FAM-labeled siRNA Control enables visualiza-tion (2–4 hours post transfection) of transfection efficiency. The human GADPH siRNA Control is a validated potent siRNA for functional evaluation of transfection efficiency.

DeliverX and DeliverX Plus siRNA Transfection Kits

U87MG cells were transfected with 10 nM of FAM-labeled siRNA Control. Two hours post transfection, bright field and fluo-rescent images of live cells were obtained.

The human brain glioblastoma astrocytoma cells were transfected with Human GADPH siRNA Control using DeliverX Plus siRNA Transfection Reagent. After 24 hours, cells were lysed and the mRNA of GADPH and cyclophilin B (internal con-trol) were quantified directly from cell lysate, without RNA purifica-tion, using QuantiGene bDNA Assay.

U87MG (human brain glioblastoma astrocytoma cells)

The differentiated 3T3-L1 mouse adipocytes were transfected with GADPH siRNA using DeliverX Plus siRNA Transfection Reagent. After 24 hours, cells were lysed and the mRNA of GADPH and cyclophilin B (internal control) were quantified directly from cell lysate, without RNA purification, using QuantiGene bDNA Assay.

Adipocytes were transfected with 10 nM of FAM-labeled siRNA Control. Two hours post transfection, bright field and fluo-rescent images of live cells were obtained.

3T3-L1 (differentiated mouse adipocytes)

Here’s how it works1. Plate cells 24 hours before

transfection.

2. Form transfection complexPrepare siRNA/Buffer-1.Prepare Transfection Reagent/Buffer-2.Combine siRNA/Buffer-1 and Transfection Reagent/Buffer-2.Incubate complex at 37°C for 20 minutes.

3. Transfect cells/quantity knockdownAdd complex, incubate 3–5 minutes at RT.Add serum-free media, incubate in 5% CO2 for 2–4 hours.

Add complete medium, incubate in 5% CO2 for 24–72 hours.

Quantify mRNA and/or protein knockdown.


Molecular Biology Products & Kits | Gene Regulation—Transfection/ Cellular ManipulationDeliverX and DeliverX Plus siRNA Transfection Kits continued...

The human THP-1 suspension cells were transfected with Human GADPH siRNA Control using DeliverX Plus siRNA Transfection Reagent. After 24 hours, cells were lysed and the mRNA of GADPH and cyclophilin B (internal control) were quantified directly from cell lysate, without RNA purification, using QuantiGene bDNA Assay.

THP-1 (human peripheral blood acute mono-cytic leukemia cells)

HeLa cells were transfected with Human GADPH siRNA Control using DeliverX siRNA Transfection Reagent. After 24 hours, cells were lysed and the mRNA of GADPH and cyclophilin B (internal control) were quantified directly from cell lysate, without RNA purification, using QuantiGene bDNA Assay.

HeLa (human cervical adenocarcinoma)

The differentiated C2C12 mouse myotubes were transfect-ed with GADPH siRNA using DeliverX Plus siRNA Transfection Reagent. After 24 hours, cells were lysed and the mRNA of GADPH and cyclophilin B (internal control) were quantified directly from cell lysate, without RNA purification, using QuantiGene® bDNA Assay.

Myotubes were transfected with 10 nM of FAM-labeled siRNA Control. Two hours post transfection, bright field and fluo-rescent images of live cells were obtained.

C2C12 (differentiated mouse myotubes)

DeliverX TAMRA-labeled Peptide Control DX1100 75 µlDeliverX Pre-formed Peptide Transfection Complex ControlDX1101 1.5 mlDeliverX Peptide Transfection Evaluation KitDX1001 0.6 ml*DeliverX Peptide Transfection KitDX1002 3.0 mlDeliverX Peptide Buffer-1DX1500 4 mlDeliverX Peptide Buffer-1DX1501 20 mlDeliverX Peptide Buffer-2DX1502 4 mlDeliverX Peptide Buffer-2DX1503 20 ml

* DeliverX Peptide Evaluation Kits are configured for new users and contain the following:

a. 0.6 ml DeliverX Peptide Transfection Reagentb. 75 µl DeliverX TAMRA-labeled Peptide Control,c. 1.5 ml DeliverX Pre-formed Peptide Transfection

Complex Control

�� Novel peptide-based, non-endosomal delivery mechanism�� Efficient delivery of a wide variety of peptides into most cell types�� Versatile—form a single transfection complex and dilute to evaluate multiple peptide concentrations�� High cell viability at optimal delivery conditions

Delivery of peptides into cells has traditionally been difficult to accomplish, both in terms of efficiency and retaining normal cellular function post transfec-tion. With the introduction of the DeliverX Peptide Transfection Kit, it is now possible to reproduce and easily deliver a wide variety of peptides into many cell types. Peptide transfection is being pursued for various applications, including modulation of cell signaling pathways for basic research, as potential therapeutic agents, and for the study of the conse-quences of pseudo up-regulation of specific cellular peptides.

Here’s how it worksDeliverX Transfection Reagents are based on novel ”MPG“ delivery technology developed at Centre de Recherches en Biochimie Macromoléculaire (CNRS) in Montpellier (France) in the laboratory of Dr. F. Heitz and Dr. G. Divita.

MPG technology uses virus-derived amphipathic peptides that directly interact with nucleic acid cargos to form nanoparticles (150-200 nm) capable of diffusing through plasma membranes and releas-ing their contents inside the cell. The mechanism of entry is receptor-independent, involves MPG/lipid membrane interactions, and avoids the endocytic pathway, thereby preventing endosomal or lysosomal degradation of cargos.

DeliverX Peptide Transfection Kits

1. MPG peptide interacts with peptide to form nanoparticle.

2. MPG/peptide nanoparticle interacts with the external side of the cellular membrane.

3. The MPG/peptide particle inserts into the membrane (a pore-like structure is created).

4. MPG/peptide particle is internalized and the peptide is decoupled from the complex.



Molecular Biology Products & Kits | Cellular Manipulation

Transfection of FITC-labeled Cdk2 Peptide Inhibitor

Delivery of FITC-labeled Cdk2 peptide inhibitor. FITC-labeled Cdk2 peptide inhibitor was transfected into HeLa cells using DeliverX Peptide Transfection Reagent.

MPG peptides can be designed to accommodate specific molecular cargos including siRNAs, single and double-strand oligonucleotides, small peptides, morpholino antisense oligonucleotides, and plasmids.

Inhibition of cell proliferationCyclin dependent kinase (Cdk) 2 is required at mul-tiple stages for progression through the mammalian cell cycle. In association with cyclin E, Cdk2 pro-motes the transition from G1 to S phase, and Cdk2/cyclinA complexes drive cells through S phase. The activities of Cdks are highly regulated by association with cyclins, whose temporal expression throughout the cell cycle is tightly controlled by phosphorylation or dephosphorylation by kinases and phosphatases, and through the actions of inhibitory proteins such as Ink4 and Cip/Kip. These key regulators of mammalian cell proliferation are excellent targets for anticancer agents. Here we demosnstrate that cellular proliferation can be inhibited by transfection of a Cdk2 inhibitory peptide into HeLa cells using the DeliverX Peptide Transfection Reagent.

Easy-to-use kitsDeliverX Peptide Transfection KitsOur DeliverX Peptide Transfection Kits are suitable for transfection of most cell types. DeliverX Peptide Transfection Kits contain the reagents required to efficiently transfect peptides into most cell types with minimal cell damage and good reproducibility when following the optimization guidelines provided in the user manual.

Transfection proficiency controlsTransfection proficiency controls are tools to enable users to optimize transfections under controlled conditions. These controls are available separately or as part of our evaluation kit.

DeliverX TAMRA-labeled Peptide Control enables visualization of transfection efficiency 1–4 hours post transfection. This validated peptide control is a 15 amino acid peptide with the TAMRA fluorophore at the carboxyl terminus. The quantity provided is sufficient for ½ of one 96-well plate.

DeliverX Pre-formed Peptide Transfection Complex Control—Enables you to test whether the DeliverX Peptide Transfection Reagent is compat-ible with your cell line or cell type. The Peptide Trans-fection Complex Control consists of the pre-formed complex of DeliverX Peptide Transfection Reagent and the TAMRA-labeled Peptide Control. The quan-tity provided is sufficient for ½ of a 96-well plate.

DeliverX Peptide Transfection Kits continued...

Cell only, time = 48 hours

DeliverX Peptide Transfection Reagent + Nega-tive Control Peptide, time = 48 hours

DeliverX Peptide Transfection Reagent + Cdk2 Peptide Inhibitor, time = 48 hours

Delivery of a Cdk2 peptide inhibitor to inhibit cell-cycle progression. HeLa cells grown in a 96-well microplate were transfected with a Cdk2 peptide inhibitor using DeliverX Peptide Transfection Reagent and incubated for 48 hours.


Molecular Biology Products & Kits | Cellular Manipulation

Cancer Cell Isolation Kit (2)CI0002 2 assaysCancer Cell Isolation Kit (4)CI0004 4 assaysCancer Cell Isolation Kit (10)CI0010 10 assays

�� Purify cancer cells from fresh biopsy samples�� Establish primary cell lines or use for biochemical assays�� Ideal for proteomics applications

The Cancer Cell Isolation Kit provides a simple, cost-effective method for isolating cancer cells from biopsy samples. Cancer cells purified using this technique can be used immediately for a wide range of applications, including biomarker identification, microarray hybridization, and genetic analysis. More-over, the whole cell isolation procedure is so gentle that cells can be used to establish primary cell lines.

Purify any population of cancer cells The ability to obtain uncontaminated cancer cell samples is one of the major bottlenecks in the study of tumor development and cancer biology. Tumor biopsies from cancer patients and animal tumor models often contain a heterogeneous population of cells that include normal tissue, blood, and cancer cells. This mixed population makes diagnosis and valid experimental conclusions difficult to obtain and interpret.

Typically, cancer cells are isolated from biopsy samples via microdissection, a time-consuming technique that requires costly equipment and surgical precision. Another conventional method is immunoprecipitation with antibodies specific to each cancer cell type. Unlike these techniques, the Cancer Cell Isolation Kit requires no specialized skill set and no expensive equipment, and can be used to isolate many cancer cell types.

Simple, cost-effective techniqueThe cancer cell isolation method only takes a couple of hours, and involves just a few rounds of incuba-tion, washing, and centrifugation. Using the same amount of starting biopsy material required for conventional methods, this technique can be used to isolate a much greater quantity of cancer cells.

Cancer cells purified using the Cancer Cell Isolation Kit can be used directly for proteomics applications such as identification of biomarkers or for RNA-based applications such as microarray hybridization and genetic analysis. Cells can also be used to purify proteins to test antibody-based cancer therapies. Alternatively, cells can be used to establish primary cell lines.

To prepare nuclear extracts from isolated cancer cells, please see the Nuclear Extraction Kit, PN AY2002 on page 102.

Kit components:Tumor cell suspension solutionTumor cell digestion solutionTumor cell purification solution100 µm filter

Cancer Cell Isolation Kit

Here’s how it works



Molecular Biology Products & Kits | Cellular Manipulation/Nuclear Extraction

Cancer Cell Isolation Kit continued...

AY2002 * *20 rxn from cell culture; 10 rxn from whole tissue

The Nuclear Extraction Kit is ideal for preparing nuclear extracts from cell culture or tissues for use with Protein/DNA Arrays or electrophoretic mobility shift assays (EMSA). This kit facilitates the extraction of functional crude nuclear proteins from various cell types or whole tissue.

Here’s how it worksThe protocol is optimized for approximately 107 cells (near-confluent 100 mm plate) or 0.5 g of tissue. First, cells are allowed to swell with hypotonic buffer. Then, the cells are disrupted, nuclei are collected, and nuclear proteins released with a high-salt buffer.

Kit components:Buffer ABuffer B100 mM DTTProtease inhibitorPhosphatase inhibitor IPhosphatase inhibitor II

Nuclear Extraction Kit


EMSA KitAY1XXX kitEMSA TF Probe SetsAY1XXXP* eaEMSA Combo Kit (any three probes)AY9999 kitEMSA Kit without Probe Set AY1000 kit*Use 4 digits from EMSA Prod. No.See chart on following page for full listing of EMSA kits.

EMSA (Electrophoretic Mobility Shift Assay) Kits* are useful tools for validating results from Protein/DNA Arrays to identify proteins that interact with DNA. You can also use them to assess the DNA-binding activity of a specific transcription factor. This rapid technique is based on the different electrophoretic mobilities of free DNA and protein/DNA complexes in native (non-denaturing) polyacrylamide gels.

Here’s how it works1. Incubate the biotin-labeled DNA probe with

your nuclear extracts which should contain your protein(s) of interest.

2. Separate the mixture on a nondenaturing polyacrylamide gel.

3. Transfer the protein/DNA complex to a nylon membrane. Incubate the blot with streptavidin-HRP and then add chemiluminescent substrate.

4. Visualize shifted bands using film or chemiluminescence imaging system that correspond to the protein DNA complexes.

To confirm whether the DNA-binding activity of a particular protein is specific, you simply add an excess of cold, unlabeled DNA probe to the reaction mixture. If binding is specific, the cold probe com-petes with the labeled DNA probe for binding to the protein. As a result, the shifted band is eliminated.

EMSA probe setsIn addition to our EMSA Kits, we offer individual EMSA probes for purchase without the other EMSA Kit components. EMSA TF Probe Sets are intended for use with our EMSA Kits. Each TF probe set is pro-vided as 25 µl (10 ng/µl) of biotin-labeled TF probe accompanied by 25 µl (330 ng/µl) of cold probe, its unlabeled form.

EMSA nomenclature For some Transcription Factors, multiple consensus sequences for the same TF have been published in the literature and, where appropriate, we have provided those variations available in our EMSA kits and arrays. For the TF’s with multiple consensus sequences, they have been denoted with numbers in parenthesis, i.e. (1), (2), (3) etc. Also, PR = Promoter and BP = Binding Protein. To obtain the consensus sequences and references for the TFs used in our arrays, please contact our Technical Support group.

Kit components:Biotin-labeled and cold TF probesBiotin-labeled and cold control probesControl nuclear extractBinding and detection reagentsEach EMSA Kit is supplied with enough reagents to run 25 reactions.

See chart on the following page for a full listing of EMSA kits.

To prepare nuclear extracts for this procedure, please see the Nuclear Extraction Kit, PN AY2002 on page 102.

EMSA (Gel-Shift) Kits and Probe Sets

PCR Methods

Molecular Biology Products & Kits | Gene Regulation— PCR Methods/Transcription Factors


Affymetrix offers a portfolio of USB branded products using PCR methods to study or quantify gene regulation events. Full descriptions of these products can be found in the PCR Tools or RNA Analysis sections of the catalog. Below is a brief summary:

Product Description

VeriQuest qPCR and qRT-PCR Master Mixes Our newest family of hot start qPCR master mixes, the VeriQuest product line, uses our own chemically modified Taq polymerase in specially formulated ready-to-use master mixes. If you’re looking for PCR efficiency and robustness which exceed the current gold standards, these mixes are for you. See pages 26-44.

HotStart-IT qPCR and qRT-PCR Master Mixes Our Hotstart-IT master mixes use a novel, patented primer sequestration method for hot start qPCR and qRT-PCR. If you’re looking for a qPCR mix with faster, easier enzyme activation, give this product a try. See pages 26-44.

Promoter Methylation PCR Kit Epigenetic analysis of your favorite promoter(s) made easy with a non-bisulfite conversion methodology. See page 24.

Poly(A) Tail-Length Assay Kit Did you know that polyadenylation length impacts the fate and function of an mRNA? Here’s an innovative kit to measure poly(A) tail-length which is far easier than the Northern blot approach. See page 158.


Molecular Biology Products & Kits | Gene Regulation—Transcription Factors

EMSA (Gel-Shift) Kits and Probe Sets continued...

AAF AY1259

ABF-1 AY1260

ACF AY1261

ACP BP AY1262

ADD-1 AY1158

ADR-1 AY1059

AF-1 AY1159

AFP-1 AY1160

Afxh/Foxo-4 AY1087

AhR/Arnt AY1060

AIC/CBF AY1161

AIC-2/3/4 AY1263

ALF-1 AY1264

α PAL AY1265

AML-1 AY1162

ANG/IRE AY1061

Antioxidant RE AY1062

AP-1 (1) AY1001

AP-1 (2) AY1055

AP-2 (1) AY1002

AP-2 (2) AY1056

AP-2 (3) AY1266

AP-2/YY1 AY1163

AP-3 (1) AY1063

AP-3 (2) AY1267

AP-4 (1) AY1064

AP-4 (2) AY1268

apo A1 PR AY1269

AR AY1003

AREB-6 AY1164

ARP AY1165

ATF (1) AY1270

ATF (2) AY1167

ATF (3) AY1271

ATF/CRE AY1166

beta-M-globin

factor B1 AY1272

beta-RE AY1065

Brn-3 AY1004

BZP AY1273

CACC AY1168

CBF (1) AY1006

CBF (2) AY1279

CBFB AY1278

CCAAT AY1169

CCAC AY1170

CD28RC (1) AY1171

CD28RC (2) AY1172

CDP AY1007

CdxA/NKX2 AY1156

CEA AY1174

C/EBP AY1005

C/EBPαγ AY1274

CEBPα(1) AY1275

CEBPα(2) AY1276

C/EBPγ AY1277

CEF-1 AY1066

CEF-2 AY1067

CETP/CRE AY1068

c-Ets-1 AY1280

c-fos BP AY1281

c-Myb (1) AY1008

c-Myb (2) AY1282

c-Myc AY1283

COUP-TF (1) AY1069

COUP-TF (2) AY1284

CP-1 AY1286

CP1/CTF/CBTF AY1175

CP-1B AY1285

CP-2 AY1287

CPE AY1176

CRE AY1289

CREB (1) AY1009

CREB (2) AY1288

CREB-2 AY1177

CREB-BP-1 AY1070

c-Rel AY1071

CSBP AY1178

CTCF AY1179

CYP1A1 AY1290

DE-1 AY1291

Dioxin Receptor AY1292

DR-0 AY1073

DR-1 AY1074

DR-2 AY1075

DR-3 AY1076

DR-4 AY1077

DR-5 AY1078

E12 (1) AY1180

E12 (2) AY1295

E12 (3) AY1293

E12/E47 AY1294

E2 AY1297

E2F-1 (1) AY1010

E2F-1 (2) AY1296

E-47 AY1079

E4BP-4 (1) AY1080

E4BP-4 (2) AY1298

E4F/ ATF AY1081

EBP-40/45 AY1181

EBP-80 AY1299

EBP-C2/NF-

muE3/YEB3 AY1300

EGF BP AY1301

EGR-1 (1) AY1011

EGR-1 (2) AY1303

EGR-1 (3) AY1302

EGR-2 (1) AY1304

EGR-2 (2) AY1305

EIL-1/2/3 AY1306

EKLF (1) AY1182

EKLF (2) AY1183

ELF AY1184

Elk-1 AY1082

ER AY1012

ETF AY1307

Ets AY1013

Ets-1/PEA-3 AY1014

ETV4/E1A-F AY1308

EVI-1 AY1083

FAST-1 AY1015

FKHR (1) AY1085

FKHR (2) AY1086

Freac-2 (1) AY1088

Freac-2 (2) AY1084

Freac-4 AY1089

Freac-7 AY1090

GAG AY1091

GAS AY1092

GAS/ISRE AY1016

Gastrin EGF RE AY1310

GATA AY1017

GATA-1 (1) AY1093

GATA-1 (2) AY1311

GATA-1/2 AY1094

GATA-2 AY1095

GATA-3 AY1096

GATA-4 AY1097

GATA-6 AY1098

GBF-1/2/3/HY5 AY1312

GCF AY1313

GFI-1 AY1099

GKLF AY1314

GR/PR AY1018

H1TF-2 AY1315

H4TF AY1185

H4TF-1 AY1316

HAS AY1108

HBS (1) AY1100

HBS (2) AY1157

HEN-1 AY1186

HFH-1 (1) AY1102

HFH-1 (2) AY1318

HFH-11β/11α AY1317

HFH-2 AY1103

HFH-3 AY1104

HFH-8 AY1105

HFH-8/3/LUN AY1187

HFS/HAS AY1101

HIF-1 AY1107

HiNF AY1188

HiNF/D3 AY1322

HiNF-A AY1319

HiNF-B/H1TF1 AY1320

HiNF-C AY1321

HLF AY1189

HMG AY1190

HNF-1 (1) AY1191

HNF-1 (2) AY1324

HNF-1 a/b/c AY1323

HNF-1α AY1192

HNF-3 (1) AY1109

HNF-3 (2) AY1110

HNF-4 (1) AY1019

HNF-4 (2) AY1111

HNF-4/COUP-TF AY1112

HNF-4α2/1 AY1325

HOX4C AY1193

HOXA-4 AY1326

HOXD-8 (1) AY1195

HOXD-8 (2) AY1196

HOXD-8/9/10 AY1194

HOXD-9/10 AY1197

HSE AY1020

HSF-2 distal AY1327

HSF-3 proximal AY1328

ICSBP AY1198

IFI-56KBP AY1330

Ikaros AY1114

IL6-RE-BP AY1331

IR-4/EBV BP AY1332

IRF-1 AY1021

IRF-1/2 AY1115

ISGF AY1333

Isl-1 AY1199

ISRE (1) AY1116

ISRE (2) AY1117

ISRE (3) AY1118

kBF-α AY1334

KKLF AY1119

KPF-1 AY1200

KTP-1 AY1201

lactoferrin BP AY1335

LCR-F1 AY1202

LEF-1 AY1203

LF-A1 (1) AY1204

LF-A1 (2) AY1336

LF-A2 AY1337

LF-B2 AY1338

LH2/Lim1 AY1339

L-III BP AY1120

LR-1 AY1205

LSF AY1340

LXRE-1 AY1341

LyF-1 (1) AY1206

LyF-1 (2) AY1207

LyF-1 (3) AY1208

MASH-1 AY1342

MAZ AY1209

MAZ (1) AY1210

MBP-1 (1) AY1343

MBP-1 (2) AY1344

mck PR E1 AY1345

mck PR E2 AY1346

MDBP (1) AY1S47

MDBP (2) AY1348

MEF-1 AY1022

MEF-2 AY1023

MEF-2(1) AY1121

MEF-2(2) AY1122

MEF-2a AYI349

MEF-3 AY1123

Mfh-1 AY1350

MHC gene PR

W Box AY1351

MRE AY1024

MSP-1 AY1124

msx-1/2/3 AY1352

hTERT-MT-Box AY1113

MTB-Zf AY1211

MTF (1) AY1212

MUSF-1 AY1125

Myb (2) AY1353

myc/CF1 AY1354

Myc-Max AY1025

MyoD AY1213

MyoG AY1214

MyTI AY1215

MZF-1 AY1126

MZF-1(1) AY1216

MZF-1(2) AY1217

MZF-1(3) AY1218

NCAM-BP AY1356

NF-1 AY1026

NF-1 (2) AY1357

NF-1/L AY1358

NF-4FA AY1219

NF-A AY1360

NF-A3 AY1359

NFAT-1 (1) AY1027

NFAT-1 (2) AY1361

NF-Atp AY1362

NF-Atx AY1363

NF-E1/YY1 AY1028

NF-E2 (1) AY1029

NF-E2 (2) AY1364

NF-E6/CP1 AY1365

NF-Gma (1) AY1220

NF-Gma (2) AY1221

NF-IL-2 AY1222

NFκB (1) AY1030

NFκB (2) AY1366

NFκB (3) AY1367

NF-Y (1) AY1127

NF-Y (2) AYI223

Nkx-2.5 AY1128

NPAS-2 AY1129

N-ras BP AY1368

NRF-1 AY1224

NRF-2 AY1225

NZF-3 AY1131

OBP AY1369

OCT (1) AY1031

OCT (2) AYI370

OCT (3) AY1371

OCT (4) AY1372

ODC AY1373

ORE AY1132

p300 AY1133

p53 (1) AY1032

p53 (2) AY1374

p53 (3) AY1226

p55 AY1227

PARP AY1134

PAX-1 AY1375

PAX-2 (1) AY1135

PAX-2 (2) AY1376

PAX-3 AY1136

PAX-4 AY1137

PAX-5 (1) AY1033

PAX-5 (2) AY1377

PAX-6 (1) AY1138

PAX-6 (2) AY1378

PAX-8 AY1139

PBGD BP AY1379

Pbx-1 AY1034

PEA3/HIP AY1228

PEBP AYI229

PEBP-2 (1) AY1230

PEBP-2 (2) AY1381

PEPCK PR GR AY1382

Pit-1(1) AY1035

Pit-1(2) AY1383

Pit-1(3) AY1384

PO-B AY1385

PPAR (1) AY1036

PPAR (2) AY1140

PPAR (3) AY1141

PPUR (1) AY1231

PPUR (2) AY1232

PRDI-BFc AY1386

PRDII-BF1 AY1387

PRE AY1037

PREB AY1388

PTF-1 AY1390

PTF-1β AY1389

PU.1 AY1391

PuF AY1233

PUR AY1392

Pur-1 AY1234

PYR AY1235

RAR/DR-5 AY1038

RAR/RXR/CF1/

MB67 AY1393

RB AY1236

RFX-1/2/3 (1) AY1237

RFX-1/2/3 (2) AY1238

RFX-1/2/3 (3) AY1394

RIPE-3a1 AY1239

RORE AY1395

RREB (1) AY1142

RREB (2) AY1143

RSRFC4 AY1144

RVF AY1396

RXR/DR-1 AY1039

SAA AY1145

SAP-1b AY1397

SIE AY1040

SIF-1 AY1240

SIF-2 AY1241

SIF-3 AY1242

Skn AY1146

SMAD/SBE AY1041

SMAD-3/4 AY1042

Snail AY1398

Sp-1 (1) AY1043

SP-1 (2) AY1243

Sp-1 (3) AY1399

SPERM-1 AY1400

SRE AY1044

SRF (1) AY1402

SRF (2) AY1401

SRF/SAP AY1244

SRY AY1147

SRY (2) AY1245

SSAP AY1403

STAT-1 AY1045

STAT-3 AY1046

STAT-4 AY1047

STAT-5 AY1048

STAT-5b AY1405

STAT-6 AY1049

Surf-2 (1) AY1406

Surf-2 (2) AY1407

T3R AY1408

Tat AY1409

Tax/CREB AY1148

TCE AY1246

TCF/LEF AY1149

TEF-1 AY1411

TEF-1/AP-5 AY1410

TFE-3 AY1247

TFE-3L AY1412

TFEB AY1413

TFIID AY1050

Tf-LF AY1248

TGT-3 AY1249

Thy-1 BP AY1414

TIF-1 AY1250

tPA BP AY1415

TR AY1051

TR/DR-4 AY1052

Transferrin BP AY1416

TREF-1/2 AY1251

TTF-1 (1) AY1417

TTF-1 (2) AY1418

TxREF/NF-lll AY1419

URE AY1420

USF-1 (1) AY1053

USF-1 (2) AY1421

VDR/DR-3 AY1054

v-Maf AY1252

v-rel 50-55K AY1422

WAP BP AY1423

WT1 (1) AY1253

WT1 (2) AY1254

WT1 (3) AY1255

X2 BP AY1256

XBP-1 (1) AY1150

XBP-1 (2) AYI424

XBP-1/X2 BP AY1257

XRF AY1151

XRE AY1425

YB-1 AY1426

Yi AY1427

ZEB AY1428

ZIA AY1152

ZIB AY1153

ZIC AY1154

ZID AY1155

Zn15 AY1429

ZNF174 AY1258

TF Prod. no. TF Prod. no. TF Prod. no. TF Prod. no. TF Prod. no. TF Prod. no. TF Prod. no.

Fra-1/JUN AY1309 HSF/HTF AY1329 myc-PRF AY1355 PCF AY1380 STAT-1/3 AY1404


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AP-1 The activator protein 1 (AP-1) transcription factor is composed of heterodimers of the Fos-activating transcription factor (ATF) and Jun subfamilies of proteins. The composition and activity of these com-plexes is regulated by the MAPK signaling pathway. AP-1 binds to transcription response elements in the promoter region of many genes involved in cell proliferation.

Lane 1. Probe Lane 2. Probe + PMA treat. HeLa nuc. ext. Lane 3. Probe + PMA treat. HeLa nuc. ext. + cold probe.

EREstrogen Receptor a (ERa) is activated by binding to estrogen that causes a conformational change and releases ERa from its inhibitory complex. ERa then binds to the estrogen response element of tar-get genes, activating gene transcription. This struc-tural change is also induced by phosphorylation of ERa, enabling substantial “cross-talk” between growth factor and estrogen receptor signaling.


EtsThe Ets-1 proto-oncogene is a member of the Ets family that demonstrates homology to the v-ets oncogene. Ets activity is regulated both by phosphorylation and via the formation of various transcription factor complexes. Activated Ets-1 binds to the Ets-binding motif and promotes expression of genes that control cell proliferation, angiogen-esis, apoptosis, and differentiation.

Lane 1. Probe Lane 2. Probe +HeLa nuc. ext.Lane 3. Probe + HeLa nuc. ext. + cold probe.

CREBThe transcription factor cAMP response element binding protein (CREB) is activated by phos-phorylation in response to a variety of extracel-lular signals, including growth factors, hormones, and neurotransmitters. Active CREB binds to the cAMP-responsive element, causing transcription of multiple genes.


EGR-1Early growth-response factor 1-(EGR-1) expression/activation is induced by signals that stimulate mito-genesis and differentiation, as well as by changes in the local cellular environment. EGR-1 interacts with a G+C-rich consensus-binding site and induces expression of many genes important in inflamma-tion, cell growth, apoptosis, and the pathogenesis of disease.

Lane 1. ProbeLane 2. Probe + Jurkat nuc. ext.Lane 3. Probe + Jurkat nuc. ext. + cold probe.

GR Glucocorticoid receptor (GR) is a nuclear hormone receptor. Upon glucocorticoid binding to GR, a change in receptor conformation occurs, causing it to dissociate from regulatory heat shock proteins. GR then translocates to the nucleus, where it binds to glucocorticoid response elements (GREs) to modulate expression of genes involved in metabolic, cardiovascular, immune, and behavioral functions.

Lane 1. ProbeLane 2. Probe + Dex. treat. HeLa nuc. ext.Lane 3. Probe + Dex. treat. HeLa nuc. ext. + cold probe.

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Our 12 Most Popular EMSA Kits



PPAR (1)Peroxisome proliferator activated receptors (PPARs) are regulated by small molecules such as plasticiz-ers, herbicides, unsaturated lipids, eicosanoids, fibrates, and thiazolidinediones. Phosphorylation of PPAR by MAP kinase causes increased transcrip-tional activity, whereas phosphorylation of PPAR by MAP kinases inhibits transcriptional activity.

Lane 1. ProbeLane 2. Probe + PMA treat. HeLa nuc. ext.Lane 3. Probe + PMA treat. HeLa nuc. ext. + cold probe

NFkB (1)The activity of NFkB is tightly regulated by interac-tions with IkB proteins. A variety of stimuli such as cytokines, growth factors, viruses, and environmen-tal hazards can trigger activation of IkB kinases, which phosphorylate IkB. This induces a conforma-tional change in IkB; NFkB then dissociates from the complex, translocates into the nucleus, and mediates transcriptional activation of target genes.

Lane 1. ProbeLane 2. Probe + TNFa treat. HeLa nuc. ext.Lane 3. Probe + TNFa treat. HeLa nuc. ext. + cold probe

p53The tumor suppressor p53 is maintained at low lev-els in cells. DNA damage induces phosphorylation or acetylation of p53, causing its stabilization and accumulation in the nucleus. p53 has the potential to act as a transcriptional activator or repressor, inducing expression of downstream genes that inhibit the cell cycle or induce apoptosis.

Lane 1. Probe.Lane 2. Probe + A431 nuc. ext.Lane 3. Probe + A431 nuc. ext. + cold probe

STAT-1The STAT-1 transcription factor undergoes tyrosine phosphorylation in response to IFNγ binding to its cognate receptor. This phosphorylation causes STAT-1 to dimerize and translocate to the nucleus, where it binds to the gamma interferon-activated site (GAS) promoter and induces the transcription of IFNγ-dependent genes. STAT-1 can also be phos-phorylated by numerous serine/threonine kinases, which increase transcriptional activity.

Lane 1. ProbeLane 2. Probe + A431 nuc. ext.Lane 3. Probe + A431 nuc. ext. + cold probe

HIF-1Hypoxia-inducible factors (HIFs) are activated by low-oxygen conditions. HIF proteins are extremely unstable in the presence of oxygen due to a high degradation rate by the proteasome. Hypoxia dramatically increases the half-life of HIF, which induces regulation of most genes activated by hypoxia.

Lane 1. ProbeLane 2. Probe + PMA treat. HeLa nuc. ext.Lane 3. Probe + PMA treat. HeLa nuc. ext. + cold probe

NFATNuclear factor of activated T cells (NFAT) is activated when T-cell receptors are stimulated. T-cell activation in turn activates calcineurin, the phosphatase that dephosphorylates NFA. This induces a conformational change in NFAT, which causes it to translocate to the nucleus, where it binds to NFAT response elements. NFAT target genes are involved in the regulation of immune responses, differentiation, and apoptosis.

Lane 1. ProbeLane 2. Probe + CD3 treat Jurkat nuc. ext.Lane 3. Probe +CD3 treat. Jurkat nuc. ext.+ cold probe

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Our 12 Most Popular EMSA Kits continued...



See chart on page 110 for a full listing of TF Activation ELISA Kits

�� Quantify activation of AP-1, E2F-1, EGR1, ERa, FKHR, GATA-1, GR, HIF-1a, HNF3b, MEF-2, MITF, NFkB c-Rel, NFkB p50, NFkB p52, NFkB p65, NkKB Rel-b, p53, PPARa, PPARγ, SMAD-2, SMAD-4, SP-1, and STAT-1�� Detect activation with as low as 0.5 µg nuclear extract �� Antibody and double-stranded DNA cis element give assay the best specificity and sensitivity possible�� Simple workflow: entire assay takes less than 4 hours

The TF Activation ELISA Kits are useful tools to detect and quantify transcription factor activation. The ELISA-based kits combine a fast, user-friendly format with a sensitive and specific assay. The assay detects activation with as low as 0.5 µg of cellular extract and the whole assay is completed in fewer than 4 hours.

An EMSA or gel shift assay typically requires 5 µg of nuclear extract and does not have the level of specificity of a TF ELISA which uses an antibody directed to the TF of interest. Each 96-well plate is comprised of individual strips of 8 wells suitable for low or high-throughput applications. These kits may be used to study drug potency, inhibitor or activator proteins, or to probe protein structure and function in signaling pathways.

Kit components:TF Antibody and HRP-conjugated secondary

antibodyConsensus DNATMB colorimetric reagent 96-well assay platePositive control nuclear extract or recombinant

protein

Here’s how it works:The TF ELISA kits use a proprietary, patent-pending technology developed for quantification of transcrip-tion factor activation. The entire procedure can be completed in less than 4 hours.

Three basic steps are involved:1. An oligonucleotide containing an TF consensus

binding site is immobilized on the 96-well plate.2. Activated TF from nuclear or whole-cell extracts

specifically binds to this oligonucleotide.3. The complex bound to the oligonucleotide is

detected by an antibody directed against the TF subunit. An additional secondary HRP-conjugated antibody provides sensitive colorimetric readout easily quantified by spectrophotometry.

TF Activation ELISA Kits

TF ELISA Kits are ideal for quantifying transcription factor binding to DNA.

PPARa ELISA KitThe PPARa ELISA Kit quantifies PPARa transcription factor activation.

ERa ELISA KitThe ERa ELISA Kit quantifies ER transcription factor activation.

Typical results obtained by using the PPARa ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. Cos-1: nuclear extract from Cos-1 cells incubated overnight in DMEM + 0.1% FBS, untreated. Cos-1+PPARa+RXRA: nuclear extract from Cos-1 cells (6–8 x 106 cells/dish) co-transfected over-night (16 hours) with 5 µg CMV-PPARa and 5 µg CMV-RXRA expres-sion vectors.

NFkB p50 ELISA KitThe NFkB p50 ELISA Kit quantifies NFkB p50 transcription factor activation.

Typical results obtained by using the ERa ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. MCF7: nuclear extract from MCF-7 cells grown in DMEM + 10% FBS. HeLa: nuclear extract from HeLa cells grown in DMEM + 10% FBS.

Typical results obtained by using the NFkB p50 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. 293 TNFa: nuclear extract from HEK293 cells that were incu-bated overnight (16 hours) in DMEM containing 0.1% FBS followed by treatment with 20 ng/ml TNFa for 30 minutes. 293: nuclear extract from HEK293 cells that were incubated overnight (16 hours) in DMEM containing 0.1% FBS, untreated.

NFκB p50

R2 = 0.9915

R2 = 0.9976

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

1.8

0 2 4 6 8 10Nuclear Extract (µg)

293 TNFα

293

OD

450

ERα

Nuclear Extract (µg)

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

0 2 4 6 8 10

OD

450

HeLaMCF7

R2 = 0.9825

R2 = 0.9961

PPARα

0

0.5

1

1.5

2

0 10 20Nuclear Extract (µg)

OD

450

R2 = 0.9972

R2 = 0.4603

Cos-1Cos-1+PPARα+RXRA




SP-1 ELISA KitThe SP-1 ELISA Kit quantifies SP-1 transcription factor activation.

Typical results obtained by using the SP-1 ELISA kit. Different amounts of recombinant human SP-1 protein as indicated were used for the assay.


OD

450 R2 = 0.9993

SP-1

0.00

0.50

1.00

1.50

2.00

2.50

3.00

0.00 5.00 10.00 15.00 20.00 25.00

A450

TF Activation ELISA Kits continued...

HIF-1a ELISA KitThe HIF-1a ELISA Kit quantifies HIF-1a transcription factor activation.

Typical results obtained by using the HIF-1a ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. Cos CoCl2: nuclear extract from Cos-1 cells treated overnight in DMEM + 0.1% FBS with 0.15 mM cobalt chloride. Cos: nuclear extract from Cos-1 cells incubated overnight in DMEM + 0.1% FBS, untreated.

p53 ELISA KitThe p53 ELISA Kit quantifies p53 transcription factor activation.

Typical results obtained by using the p53 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. MCF7: nuclear extract from MCF-7 cells incubated overnight in DMEM + 0.1% FBS, untreated. MCF7 H2O2: nuclear extract from MCF-7 cells incubated overnight in DMEM + 0.1% FBS, then treated with 0.2 mM H2O2 for 3 hours.

HIF-1α


OD

450

R2 = 0.9986

R2 = 0.9876

Cos CoCl2Cos

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

0 2 4 6 8 10

Nuclear Extract (µg)O

D 4

50

R2 = 0.9974

R2 = 0.9969

p53

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

0 2 4 6 8 10

MCF7

MCF7 H2O2

AP-1 ELISA KitThe AP-1 ELISA Kit quantifies AP-1 transcription factor activation.

GR ELISA KitThe GR ELISA Kit quantifies GR transcription factor activation.

Typical results obtained by using the AP-1 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. 293 PMA: nuclear extract from HEK293 cells that were incubated over-night (16 hours) in DMEM containing 0.1% FBS, then treated with 10 ng/ml PMA for 4 hours. 293: nuclear extract from HEK293 cells that were incubated overnight (16 hours) in DMEM containing no FBS, untreated.

Typical results obtained by using the GR ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. HeLa: nuclear extract from HeLa cells that were incubated overnight (16 hours) in DMEM + 0.1% FBS, untreated. HeLa Dex: nuclear extract from HeLa cells incubated overnight in DMEM + 0.1% FBS followed by treatment with 100 nM dexamethasone for 1 hour.


OD

450

R2 = 0.9414

R2 = 0.9957

AP-1

293

293 PMA

0.0

0.2

0.4

0.6

0.8

1.0

1.2

0 2 4 6 8 10


OD

450

R2 = 0.9899

R2 = 0.9865

GR

HeLa Dex

HeLa

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

0 2 4 6 8 10

STAT-1 ELISA KitThe STAT-1 ELISA Kit quantifies transcription factor activation, specifically that of STAT-1.

Typical results obtained by using the STAT-1 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. HeLa: nuclear extract from HeLa cells that were incubated overnight (16 hours) in DMEM containing no FBS, untreated. HeLa IFNγ: nuclear extract from HeLa cells incubated overnight in DMEM + 0.1% FBS followed by treatment with 5 ng/ml IFNγ for 2 hours.


OD

450

R2 = 0.9891

R2 = 0.8107

HeLa

HeLa IFNg

STAT-1

0.0

0.2

0.4

0.6

0.8

1.0

1.2

0 2 4 6 8 10




MEF-2 ELISA KitThe MEF-2 ELISA Assay Kit quantifies MEF-2 tran-scription factor activation.

GATA-1 ELISA KitThe GATA-1 ELISA Kit quantifies transcription factor activation, specifically that of GATA-1.

Typical results obtained by using the MEF-2 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. U937: nuclear extract from U937 cells grown in DMEM + 10% FBS. C2C12: nuclear extract from C2C12 cells grown in DMEM + 10% FBS.

Typical results obtained by using the GATA-1 ELISA kit. Different amounts of recombinant human GATA-1 protein as indicat-ed were used for the assay.


OD

450

R2 = 0.9955

R2 = 0.9945

U937

C2C12

MEF-2

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

0 2 4 6 8 10Nuclear Extract (µg)

OD

450

R2 = 0.999A450

GATA-1

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

0 10 20 30 40

SMAD-2 ELISA Kit

HNF3b ELISA Kit

Typical results obtained by using the SMAD-2 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. HeLa: nuclear extract from HeLa cells incubated overnight in DMEM + 0.1% FBS, untreated. HeLa TGFb: nuclear extract from HeLa cells that were incubated overnight (16 hours) in DMEM con-taining 0.1% FBS followed by treatment with 10 ng/ml TGFb for 30 minutes.

Typical results obtained by using the HNF3b ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. HepG2: nuclear extract from HepG2 cells that were grown in DMEM containing 10% FBS. HeLa: nuclear extract from HeLa cells grown in DMEM + 10% FBS.

SMAD-4 ELISA Kits

Typical results obtained by using the SMAD-4 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. HeLa: nuclear extract from HeLa cells incubated overnight in DMEM + 0.1% FBS, untreated. HeLa TGFb: nuclear extract from HeLa cells that were incubated overnight (16 hours) in DMEM con-taining 0.1% FBS followed by treatment with 10 ng/ml TGFb for 30 minutes.


OD

450

R2 = 0.9878

R2 = 0.6832

HepG2

HeLa

HNF3β

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

0 5 10 15


OD

450

R2 = 0.9967HeLa

HeLa TGFb

SMAD-2

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

0 2 4 6 8 10

R2 = 0.9619


OD

450

HeLa

HeLa TGFb R2 = 0.9936

R2 = 0.987

SMAD-4

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0 2 4 6 8 10



Typical results obtained by using the FKHR ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. Raji: nuclear extract from Raji cells that were grown in RPMI-1640 containing 10% FBS. HeLa: nuclear extract from HeLa cells grown in DMEM + 10% FBS.

FKHR ELISA Kits


OD

450

Raji

HeLa R2 = 0.9975

R2 = 0.954

FKHR

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0 2 4 6 8 10



E2F-1 ELISA KitThe E2F-1 ELISA Kit quantifies E2F-1 transcription factor activation.

Typical results obtained by using the E2F-1 ELISA kit. Different amounts of nuclear extract as indicated were used for the assay. HeLa: nuclear extract from HeLa cells that were incubated 3 days in DMEM containing no FBS. HeLa serum: nuclear extract from HeLa cells grown in DMEM + 10% FBS.


OD

450

HeLa

HeLa serum

R2 = 0.9892

R2 = 0.9899

E2F-1

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

0 2 4 6 8 10

Product Size Prod. no.AP-1 Cold Probe 25 µl EK0504AP-1 ELISA Kit kit EK1040AP-1 ELISA Kit with Nuclear Extraction Kit kit EK1041Control HEK292 PMA Nuclear Extract 20 µl EK0505E2F-1 ELISA Kit kit EK2000E2F-1 ELISA Kit with Nuclear Extraction Kit kit EK2001ERa ELISA Kit kit EK1030ERa ELISA Kit with Nuclear Extraction Kit kit EK1031ER Control MCF7 Nuclear Extract 20 µl EK0503ERE Cold Probe 25 µl EK0502FKHR ELISA Kit kit EK1400FKHR ELISA Kit with Nuclear Extraction Kit kit EK1401GATA-1 Cold Probe 25 µl EK0520GATA-1 Control Recombinant Protein 20 µl EK0521GATA-1 ELISA Kit kit EK1100GATA-1 ELISA Kit with Nuclear Extraction Kit kit EK1101GR Cold Control Probe 25 µl EK0508GR Control Hela (DEX) Nuclear Extract 20 µl EK0509GR ELISA Kit kit EK1060GR ELISA Kit with Nuclear Extraction Kit kit EK1061HIF Cold Control Probe 25 µl EK0500HIF Control Cos1 Nuclear Extract 20 µl EK0501HIF-1a ELISA Kit kit EK1020HIF-1a ELISA Kit with Nuclear Extraction Kit kit EK1021HNF-3b ELISA Kit kit EK1800HNF-3b ELISA Kit with Nuclear Extraction Kit kit EK1801MEF-2 Cold Control Probe 25 µl EK0512MEF-2 Control Nuclear Extract 20 µl EK0513MEF-2 ELISA Kit kit EK1080MEF-2 ELISA Kit with Nuclear Extraction Kit kit EK1081NFkB c-Rel ELISA Kit kit EK4100

Product Size Prod. no.NFkB c-Rel ELISA Kit with Nuclear Extraction Kit kit EK4101NFkB Control Nuclear Extract 20 µl EK0523NFkB Cold Control Probe 25 µl EK0522NFkB p50 ELISA Kit kit EK1110NFkB p50 ELISA Kit with Nuclear Extraction Kit kit EK1111NFkB p52 ELISA Kit kit EK3100NFkB p52 ELISA Kit with Nuclear Extraction Kit kit EK3101NFkB p65 ELISA Kit kit EK1120NFkB p65 ELISA Kit with Nuclear Extraction Kit kit EK1121NFkB Rel-b ELISA Kit kit EK5100NFkB Rel-b ELISA Kit with Nuclear Extraction Kit kit EK5101p53 Cold Control Probe 25 µl EK0506p53 ELISA Kit kit EK1050p53 ELISA Kit with Nuclear Extraction Kit kit EK1051PPAR Cold Control Probe 25 µl EK0516PPARa Cold Control Probe 25 µl EK0518PPARa Control Cos1 Nuclear Extract 20 µl EK0519PPARa ELISA Kit kit EK1310PPARa ELISA Kit with Nuclear Extraction Kit kit EK1311SMAD-2 ELISA Kit kit EK1600SMAD-2 ELISA Kit with Nuclear Extraction Kit kit EK1601SMAD-4 ELISA Kit kit EK1500SMAD-4 ELISA Kit with Nuclear Extraction Kit kit EK1501SP-1 Cold Probe 25 µl EK0514SP-1 Control Recombinant Protein 20 µl EK0515SP-1 ELISA Kit kit EK1090SP-1 ELISA Kit with Nuclear Extraction Kit kit EK1091STAT-1 Cold Control Probe 25 µl EK0510STAT-1 Control MCF7 (IFNγ) 20 µl EK0511STAT-1 ELISA Kit kit EK1070STAT-1 ELISA Kit with Nuclear Extraction Kit kit EK1071

TF Activation ELISA Kits



LRXXXX 10 µgPlease order using the product number identified on the chart below.

�� Physiologically relevant—contains cloned promoter region of specific genes �� Flexible—use for in vivo assays �� Quantitative—use luciferase activity to measure promoter activity �� Safe—no radioactivity required �� Simple—easily applied to high-throughput screening

With the Gene Promoter Reporter Vectors, you can quantify the promoter activity of 34 genes. The luciferase assay is a simple, straightforward, and very effective means of measuring promoter response in cells.

The actual length of the cloned promoter region dif-fers for each Gene Promoter Reporter Vector.

Gene Promoter Reporter Vectors

The Gene Promoter Reporter Vector is available for 34 different genes. The insert contains approximately 800 bp of 5' flanking region and 50-200 bp of untranslated region of exon 1 of the human genes listed below. This ~1 kb region is located upstream of the start codon of luciferase gene (Panel A.) The vector backbone contains an ampicillin-resistance gene which allows ampicillin selec-tion in E. coli, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production (Panel B).

The Gene Promoter Reporter Vectors in HeLa cells. HeLa cells were transfected with each of the vectors listed and then treated with/without phorbol myristate acetate (PMA). The luciferase activity was significantly enhanced with PMA treatment for TNFa, RB1, and JUNB Gene Promoter Vectors, indicating that these three gene promoters are activated by PMA in HeLa cells, while others are not.

Product Size Prod. no.14-3-3-sigma (SFN) Gene Promoter Reporter Vector 10 µg LR1009ABL1 Gene Promoter Reporter Vector 10 µg LR1001APC Gene Promoter Reporter Vector 10 µg LR1019ATF2 Gene Promoter Reporter Vector 10 µg LR1007BRCA1 Gene Promoter Reporter Vector 10 µg LR1002CFTR Gene Promoter Reporter Vector 10 µg LR1020CGPR (CALCR) Gene Promoter Reporter Vector 10 µg LR1003CIITA Gene Promoter Reporter Vector 10 µg LR1010COX2 (PTGS2) Gene Promoter Reporter Vector 10 µg LR1029ERa Gene Promoter Reporter Vector 10 µg LR1017FHIT Gene Promoter Reporter Vector 10 µg LR1032G6PD Gene Promoter Reporter Vector 10 µg LR1021GAGE1 Gene Promoter Reporter Vector 10 µg LR1004hMLH1 Gene Promoter Reporter Vector 10 µg LR1022HOXA2 Gene Promoter Reporter Vector 10 µg LR1030HOXA5 Gene Promoter Reporter Vector 10 µg LR1031IFNγ Gene Promoter Reporter Vector 10 µg LR1011

Product Size Prod. no.IGRP Gene Promoter Reporter Vector 10 µg LR1012IL1a Gene Promoter Reporter Vector 10 µg LR1013IL2 Gene Promoter Reporter Vector 10 µg LR1014IL4 Gene Promoter Reporter Vector 10 µg LR1023IRF7 Gene Promoter Reporter Vector 10 µg LR1005JUNB Gene Promoter Reporter Vector 10 µg LR1024MDR1 Gene Promoter Reporter Vector 10 µg LR1028MyoD Gene Promoter Reporter Vector 10 µg LR1025POMC Gene Promoter Reporter Vector 10 µg LR1006RB1 Gene Promoter Reporter Vector 10 µg LR1018STAT1 Gene Promoter Reporter Vector 10 µg LR1008SURVIVIN Gene Promoter Reporter Vector 10 µg LR1016THBS2 Gene Promoter Reporter Vector 10 µg LR1033TIMP3 Gene Promoter Reporter Vector 10 µg LR1034TNFa Gene Promoter Reporter Vector 10 µg LR1015VHL Gene Promoter Reporter Vector 10 µg LR1026v-myc Gene Promoter Reporter Vector 10 µg LR1027

Reporter vectors

A.

B.



LRXXXX See belowPlease order using the product number identified in the chart below.

�� Quantify transactivation of transcription factor binding in vivo�� Target over 50 specific transcription factors�� Establish cell lines for cell-based assays and drug discovery

With Luciferase Reporter Vectors, you can easily measure the binding activities of transcription factors (TFs) in vivo. TF activation is measured by induced expression of firefly luciferase. This luciferase-based assay is simple, flexible, and rapid, generating quan-titative results in hours.

By simply transfecting a vector into cells, you can examine the effects of various stimuli on a given signaling pathway in vivo, such as CRE-luc for G-protein coupled receptors, NFkB-luc for TNFa, and STAT-luc for T cell antigen receptor. These vectors can be used to monitor disease-related TFs, such as PPAR for diabetes, NFkB for inflammatory disease, MEF-2 or MIF-1 for heart disease, and p53 for cancer.

Like our EMSA gel-shift kits, the Reporter Vectors are also ideal for validating the results obtained using Protein/DNA Arrays. But while EMSA is an in vitro assay, the Reporter Vectors allow you to monitor TF activity in living cells.

The Luciferase Reporter Vectors have been specially constructed to monitor transactivation of over 50 specific TFs. Each vector contains multiple repeats of a specific transcription factor binding element, or cis-element. Binding of a TF to its cis-element drives the expression of firefly luciferase. Light emitted from the chemical reaction is directly proportional to the amount of expressed luciferase, and thus the binding activity of the targeted TF.

Using Reporter Vectors is as simple as performing a standard luciferase assay, a procedure that takes just a few hours. Cells are transfected with a reporter vector, treated with corresponding stimuli, and then lysed to measure the luciferase activity.

By transfecting a reporter vector into cells along with vectors conferring resistance to hygromycin or neomycin, you can establish stable cell lines for use in cell-based assays.

Each Luciferase Reporter Vector comes with a Control Reporter Vector.

Luciferase Reporter Vectors

293, HeLa, or Cos-1 cells were transiently transfected with various Luciferase Reporter Vectors. The cells were then treated with inducers, the activity of each TF was then quantified by measur-ing luciferase activity.

Product Size Prod. no.TA-luc (Empty Control) Luciferase Reporter Vector 20 µg LR0000AP1(1) Luciferase Reporter Vector 10 µg LR0002AP1(2) Luciferase Reporter Vector 10 µg LR0003AP3 Luciferase Reporter Vector 10 µg LR0005AR Luciferase Reporter Vector 10 µg LR0007CRE(1) Luciferase Reporter Vector 10 µg LR0093E2F(1) Luciferase Reporter Vector 10 µg LR0016E2F(2) Luciferase Reporter Vector 10 µg LR0017E2F1(3) Luciferase Reporter Vector 10 µg LR0094ELK1 Luciferase Reporter Vector 10 µg LR0019ER Luciferase Reporter Vector 10 µg LR0020GAS / ISRE Luciferase Reporter Vector 10 µg LR0026GAS Luciferase Reporter Vector 10 µg LR0025GATA Luciferase Reporter Vector 10 µg LR0027GATA1 / GATA2 Luciferase Reporter Vector 10 µg LR0029GATA1 Luciferase Reporter Vector 10 µg LR0028GATA2 Luciferase Reporter Vector 10 µg LR0030GATA3 Luciferase Reporter Vector 10 µg LR0031GATA4 Luciferase Reporter Vector 10 µg LR0032GR Luciferase Reporter Vector 10 µg LR0033HIF1 Luciferase Reporter Vector 10 µg LR0128HSE Luciferase Reporter Vector 10 µg LR0038IRF1 Luciferase Reporter Vector 10 µg LR0039ISRE Luciferase Reporter Vector 10 µg LR0040

Product Size Prod. no.MEF1 Luciferase Reporter Vector 10 µg LR0043MEF2 Luciferase Reporter Vector 10 µg LR0044MEF3 Luciferase Reporter Vector 10 µg LR0045NF-AT Luciferase Reporter Vector 10 µg LR0050NFkB(1) Luciferase Reporter Vector 10 µg LR0051NFkB(2) Luciferase Reporter Vector 10 µg LR0052p53 Luciferase Reporter Vector 10 µg LR0057PR Luciferase Reporter Vector 10 µg LR0067RAR Luciferase Reporter Vector 10 µg LR0068RXR Luciferase Reporter Vector 10 µg LR0070RXR(2) Luciferase Reporter Vector 10 µg LR0114Smad Luciferase Reporter Vector 10 µg LR0116Smad3 / Smad4 Luciferase Reporter Vector 10 µg LR0115Sp1 Luciferase Reporter Vector 10 µg LR0071SP1(2) Luciferase Reporter Vector 10 µg LR0117SRE Luciferase Reporter Vector 10 µg LR0072SRF Luciferase Reporter Vector 10 µg LR0073STAT1 p84/p91 Luciferase Reporter Vector 10 µg LR0075STAT1(2) Luciferase Reporter Vector 10 µg LR0127STAT3(1) Luciferase Reporter Vector 10 µg LR0077STAT4 Luciferase Reporter Vector 10 µg LR0079VDR Luciferase Reporter Vector 10 µg LR0087YY1 Luciferase Reporter Vector 10 µg LR0090

Reporter vectors



The Luciferase Reporter Vectors can be used to quantify induction in various cell lines. For the data shown above, 293, HeLa, and Cos-1 cells were transiently transfected with the indicated Luciferase Reporter Vector, in the presence or absence of an inducer molecule. The fold-induction was calculated for each vector relative to the unstimulated control. No induction by tested inducers was observed with the Negative Control Vectors.

Luciferase Reporter Vectors continued...



MEXXXX 15 µgPlease order using the product number identified on the chart below.

�� Ready for transfection into cells for gene expression studies �� Fully-sequenced TF cDNA inserts

Transcription factor (TF) cDNA sequences are identi-cal to those on NCBI Refseq.

Mammalian Full-length TF Expression Vectors

Cloned human TF proteins are expressed at high levels from the constitutive CMV promoter. The SV40 polyA sequence ensures efficient processing of the 3' end of the mRNAs. The vector backbone contains an SV40 origin for replication in mammalian cells. A neomycin-resistance gene allows kanamycin selection in E. coli and neomycin selection in eukaryotic cells. The vector backbone also contains a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.

pCMV-NFkB/p505748 bp

pUC ori

SV40 poly A

Not I

pSV40

Kan/Neor

NfkB/p50

pCMV

f1 ori

Xho I

HSV TK poly A

Product Size Prod. no.ATF1 CMV Expression Vector 15 µg ME0059ATF3 CMV Expression Vector 15 µg ME0055BTG2 CMV Expression Vector 15 µg ME0065CDX2 CMV Expression Vector 15 µg ME0060CITED1 CMV Expression Vector 15 µg ME0020E2F2 CMV Expression Vector 15 µg ME0010ERa CMV Expression Vector 15 µg ME0043FOS CMV Expression Vector 15 µg ME0044GATA1 CMV Expression Vector 15 µg ME0049GFI1 CMV Expression Vector 15 µg ME0066GR (NR3C1) CMV Expression Vector 15 µg ME0007HNF4γ CMV Expression Vector 15 µg ME0062IRF1 CMV Expression Vector 15 µg ME0014JUN CMV Expression Vector 15 µg ME0045MAFK CMV Expression Vector 15 µg ME0018MAX CMV Expression Vector 15 µg ME0019MYC CMV Expression Vector 15 µg ME0052 NF-ATC1 CMV Expression Vector 15 µg ME0046

Product Size Prod. no.NFkB p50 CMV Expression Vector 15 µg ME0005NFkB p65 (RelA) CMV Expression Vector 15 µg ME0006NF-YC CMV Expression Vector 15 µg ME0022NR0B1 CMV Expression Vector 15 µg ME0061NR1I2 CMV Expression Vector 15 µg ME0024NR1I3 CMV Expression Vector 15 µg ME0025p53 CMV Expression Vector 15 µg ME0047PBX1 CMV Expression Vector 15 µg ME0054PPARa CMV Expression Vector 15 µg ME0001PPARb (PPARD) CMV Expression Vector 15 µg ME0002PPARγ-1 CMV Expression Vector 15 µg ME0003PPARγ-2 CMV Expression Vector 15 µg ME0004PREB CMV Expression Vector 15 µg ME0029SMAD1 (MADH1) CMV Expression Vector 15 µg ME0071SRY CMV Expression Vector 15 µg ME0032STAT1 CMV Expression Vector 15 µg ME0033STAT5B CMV Expression Vector 15 µg ME0035USF1 CMV Expression Vector 15 µg ME0040

Expression vectors



cAMP/Calcium Protein/DNA ArrayMA1500 kitcAMP/Calcium Protein/DNA Array RefillMA1600 kitCell Growth Protein/DNA ArrayMA1505 kitCell Growth Protein/DNA Array RefillMA1605 kitNuclear Receptor Protein/DNA ArrayMA1510 kitNuclear Receptor Protein/DNA Array RefillMA1610 kit

�� Focus on transcription factors involved in specific biological processes

We offer the Function-Specific Protein/DNA Arrays for function-specific analysis of eukaryotic TFs, allowing you to focus on factors involved in specific biological processes.

cAMP/Calcium Protein/DNA ArrayWith the cAMP/Calcium Protein/DNA Array, you can easily determine how the activities of cAMP- and calcium-regulated TFs change under different condi-tions. The array enables you to survey the activity of 20 TFs that are known to be regulated by cAMP and calcium.

Nuclear Receptor Protein/DNA ArrayThe Nuclear Receptor Protein/DNA Array is designed for researchers who are interested in how the activities of the lipophilic nuclear hormone receptor superfamily receptors—including steroid and thyroid hormones, active forms of vitamin A (retinoids), and

vitamin D—change under different conditions. With our Nuclear Receptor Protein/DNA Array, you can profile the activities of 15 nuclear receptors.

Cell Growth Protein/DNA ArrayThe Cell Growth Protein/DNA Array provides a good starting point for researchers who are looking to map the series of events leading from extracellular stimuli to gene transcription controlling growth and differen-tiation. This array allows screening for 20 TFs that are key players in cell growth and differentiation.

Kit components:Three array membranesHybridization and wash solutionsSpin column and buffersHRP chemiluminescent reagentsControl nuclear extract

Function-Specific Protein/DNA Arrays

TF-TF Interaction Array I*MA5010 kit TF-TF Interaction I RefillMA5110 kitTF-TF Interaction Array II*MA5011 kitTF-TF Interaction II RefillMA5111 kitTF-TF Interaction Array III*MA5012 kitTF-TF Interaction III RefillMA5112 kit

* See Protein/DNA Array chart on page 117 for cis-elements included on TF Interaction Array I, II and III.

�� Identify interactions between different transcription factors (TFs)�� Analyze transcription factor complex formation following different stimuli�� Monitor potential cross-talk between intracellular signaling cascades

TF-TF Interaction Arrays enable you to determine how a particular TF interacts with multiple other TFs—all in one experiment. Unlike traditional methods, such as co-immunoprecipitation and super-gel shift, this array-based method generates a global view of TF-TF interactions. The nonradioactive, all-in-one system offers the high sensitivity of HRP-based chemiluminescence detection.

Each consensus sequence immobilized on our TF-TF Interaction Array corresponds to a specific DNA-binding TF. We have 3 different arrays profiling a selection of transcription factors.


Here’s how it worksThe TF-TF Interaction Array, instead of directly measuring interactions between proteins, employs an exquisitely sensitive indirect approach. This technology is based on the fact that every TF binds to a specific DNA cis-element. The proteins present in a TF complex can be determined by isolating a complex by immunoprecipitation and monitoring which cis-elements bind to this complex—indicating which TF are present. In this way, you can quickly see how one TF interacts with multiple other TFs. Individual interactions can then be confirmed by co-immunoprecipitation or super-gel shift.

TF-TF Interaction Arrays

A. A431

The TF-TF Interaction Array I profiles interactions between multiple transcription factors. Nuclear extracts from A431 and Saos cells were incubated with Probe Mix I. The TF of interest, p53, was then immunoprecipitated using a p53-specific Ab, and the pro-tein-bound probes were isolated and hybridized to the TF-TF Interaction Array I revealing p53-specific interactions in A431 (Panel A), and no interactions in the p53 negative Saos2 cell line (Panel B).

B. Saos2



Protein/DNA Array IMA1210 kitProtein/DNA Array I Refill KitMA1310 kitProtein/DNA Array IIMA1211 kitProtein/DNA Array II Refill KitMA1311 kitProtein/DNA Array IIIMA1212 kitProtein/DNA Array III Refill KitMA1312 kitProtein/DNA Array IVMA1213 kitProtein/DNA Array IV Refill KitMA1313 kitProtein/DNA Array VMA1214 kitProtein/DNA Array V Refill KitMA1314 kitCombo Protein/DNA ArrayMA1215 kitCombo Protein/DNA Refill KitMA1315 kit

�� Profile the activities of multiple transcription factors at once�� Monitor cross-talk between intracellular signaling cascades�� Characterize promoter regions of novel genes�� Determine, by high-throughput analysis, novel mechanisms of transcriptional regulation�� Greatly improved probe recovery system increases assay sensitivity

Protein/DNA Arrays simplify the functional analysis of eukaryotic transcription factors (TFs). Our array-based technology is a significant improvement over cumbersome gel mobility-shift assays, which permit the characterization of only a single TF at a time. With the Protein/DNA Arrays, you can profile the activities of multiple TFs simultaneously. The Protein/DNA Arrays can be used to study TF activation in a variety of biological processes, including cell proliferation, differentiation, transformation, and apoptosis.

Five versions of the Protein/DNA Array are currently available. All five arrays are spotted with unique, non-overlapping consensus-binding sequences corresponding to specific TFs, which were carefully selected after a survey of the literature. For your convenience we also offer a Combo version of the Protein/DNA Array spotted with all 345 cis-elements.


The new filter-based spin column system is a gel-free separation which makes probe isolation and recovery a quick and simple three-step process. This spin column separation procedure significantly increases the amount of probe recovered, providing a more sensitive assay that requires much less starting material. The reduced sample handling also helps minimize variability between experimental samples.

With increased probe recovery and ease of use, this system enables the whole experiment, from start to finish, to be completed in one and a half days. The added ease of sample preparation allows simultane-ous analysis of many more samples.

Kit components:Three array membranesHybridization and wash solutionsSpin column reagentsChemiluminescence reagentsControl nuclear extract

Refill Kits for reprobing array membranes can be purchased separately.

References:1. Jing, H. et al. (2004) J. Biol. Chem. 279,

55176–55186.2. Peterson, R. S. et al. (2004) J. Cell Biol. 166,

1081–1091.3. Barnhart, B. C. et al. (2004) EMBO J. 23,

3175–3185.4. Govindarajan, B. et al. (2003) J. Biol. Chem. 278,

9790–9795.

For some Transcription Factors, mutliple consensus sequences for the same TF have been published in the literature and, where appropriate, we have provided those variations available in our EMSA kits and arrays. For the TF’s with mutliple consensus sequences, they have been denoted with numbers in parenthesis, i.e. (1), (2), (3) etc. To obtain the consen-sus sequences and references for the TFs used in our arrays, please contact our Technical Support group.

Related products

Looking to profile transcription factors from a larger number of samples? Call our Technical Support team and ask about the Procarta Transcription Factor Plex Assays.

Protein/DNA Arrays

1. Mix biotin-labeled DNA binding probes with nuclear extract or cell lysate, this allows the formation of “DNA/Protein” complexes.

2. After incubation, these complexes are added to a spin col-umn which allows the separation of unbound probes.

3. After elution of the DNA/Protein com-plexes, they are denatured to liberate the free probe.

4. Hybridize the free probe to the mem-brane, which con-tains an array of Transcription Factor consensus binding sequences.


The Protein/DNA Arrays simultaneously detect changes in activities of multiple transcription factors. The array data was obtained using the Protein/DNA Array I. Nuclear extracts from untreated (panel A) or TNFa treated (panel B) HeLa cells were incubated with probe mix. The protein bound Probes were then isolated and hybridized to the Protein/DNA Array I membrane. Chemiluminescent images were collected using the FluorChem™ Imager (Alpha Innotech) revealing changes in protein binding activities of AP-1 and NFkB.

A. Untreated HeLa cells

B. TNFa treated HeLa cells



Protein/DNA Arrays continued...

ADR-1

Afxh/Foxo-4

AhR/Arnt

ANG/IRE

Antioxidant RE

AP-3 (1)

AP-4 (1)

beta-RE

CdxA/NKX2

CEF-1

CEF-2

CETP/CRE

COUP-TF (1)

CREB-BP-1

c-Rel

DR-0

DR-1

DR-2

DR-3

DR-4

DR-5

E-47

E4BP-4 (1)

E4F/ ATF

Elk-1

EVI-1

FKHR (1)

FKHR (2)

Freac-2 (1)

Freac-2 (2)

Freac-4

Freac-7

GAG

GAS

GATA-1 (1)

GATA-1/2

GATA-2

GATA-3

GATA-4

GATA-6

GFI-1

HAS

HBS (1)

HBS (2)

HFH-1 (1)

HFH-2

HFH-3

HFH-8

HFS/HAS

HIF-1

HNF-3 (1)

HNF-3 (2)

HNF-4 (2)

HNF-4/COUP-TF

Ikaros

IRF-1/2

ISRE (1)

ISRE (2)

ISRE (3)

KKLF

L-III BP

MEF-2 (2)

MEF-2 (3)

MEF-3

MSP-1

MT-Box

MUSF-1

MZF-1 (1)

NF-Y (1)

Nkx-2.5

NPAS-2

NZF-3

ORE

p300

PARP

PAX-2 (1)

PAX-3

PAX-4

PAX-6 (1)

PAX-8

PPAR (2)

PPAR (3)

RREB (1)

RREB (2)

RSRFC4

SAA

Skn

SRY (1)

Tax/CREB

TCF/LEF

XBP-1 (1)

XRE (1)

ZIA

ZIB

ZIC

ZID

Protein/DNA Array II, TF-TF Interaction Array II, TF-Specific-TF Arrays II

ADD-1

AF-1

AFP-1

AIC/CBF

AML-1

AP-2/YY1

AREB-6

ARP

ATF (2)

ATF/CRE

CACC

CCAAT

CCAC

CD28RC (1)

CD28RC (2)

Cdx-2

CEA

CP1/CTF/CBTF

CPE

CREB-2

CSBP

CTCF

E12 (1)

EBP-40/45

EKLF (1)

EKLF (2)

ELF

H4TF

HEN-1

HFH-8/3/LUN

HiNF

HLF

HMG

HNF-1 (1)

HNF-1α

HOX4C

HOXD-8 (1)

HOXD-8 (2)

HOXD-8/9/10

HOXD-9/10

ICSBP

Isl-1

KPF-1

KTP-1

LCR-F1

LEF-1

LF-A1 (1)

LR-1

LyF-1 (1)

LyF-1 (2)

LyF-1 (3)

MAZ (1)

MAZ (2)

MTB-Zf

MTF (1)

MyoD

MyoG

MyTI

MZF-1 (2)

MZF-1 (3)

MZF-1 (4)

NF-4FA

NF-Gma (1)

NF-Gma (2)

NF-IL-2

NF-Y (2)

NRF-1

NRF-2

p53 (2)

p55

PEA3/HIP

PEBP

PEBP-2 (1)

PPUR (1)

PPUR (2)

PuF

Pur-1

PYR

RB

RFX-1/2/3 (1)

RFX-1/2/3 (2)

RIPE-3a1

SIF-1

SIF-2

SIF-3

SP-1 (2)

SRF/SAP

SRY (2)

TCE

TFE-3

Tf-LF

TGT-3

TIF-1

TREF-1/2

v-Maf

WT1 (1)

WT1 (2)

WT1 (3)

X2 BP

XBP-1/X2 BP

ZNF174

Protein/DNA Array III, TF-TF Interaction Array III, TF-Specific-TF Arrays III

AAF

ABF-1

ACF

ACP BP

AIC-2/3/4

ALF-1

α PAL

AP-2 (3)

AP-3 (2)

AP-4 (2)

apo A1 PR

ATF (1)

ATF (3)

beta-M-globin

factor B1

BZP

CBF (2)

CBFB

CEBP (2)

CEBP (3)

CEBP (4)

CEBP (5)

c-Ets-1

c-fos BP

c-Myb (2)

c-Myc

COUP-TF (2)

CP-1

CP-1B

CREB (2)

CYP1A1

DE-1

Dioxin Receptor

E12 (3)

E12/E47

E2

E2F-1 (2)

E4BP-4 (2)

EBP-80

EBP-C2/

NF-muE3/YEB3

EGF BP

EGR-1 (2)

EGR-2 (1)

EIL-1/2/3

ETF

ETV4/E1A-F

Fra-1/JUN

Gastrin EGF RE

GATA-1 (2)

GBF-1/2/3/HY5

GKLF

H4TF-1

HFH-1 (2)

HFH-11β/11α

HiNF/D3

HiNF-A

HiNF-B/H1TF1

HNF-1 (2)

HNF-1 a/b/c

HNF-4α2/1

HOXA-4

HSF/HTF

HSF-2 distal

HSF-3 proximal

IFI-56KBP

IL6-RE-BP

IR-4/EBV BP

ISGF

kBF-α

lactoferrin BP

LF-A1 (2)

LF-A2

LF-B2

LH2/Lim1

LSF

LXRE-1

MASH-1

MBP-1 (1)

MBP-1 (2)

Protein/DNA Array IV

mck PR E1

MDBP (1)

MDBP (2)

MEF-2a

Mfh-1

MHC gene PR

W Box

msx-1/2/3

Myb (2)

myc/CF1

myc-PRF

NCAM-BP

NF-1 (2)

NF-1/L

NF-A

NF-A3

NFAT-1 (2)

NF-Atp

NF-Atx

NF-E2 (2)

NF-E6/CP1

NFκB (2)

NFκB (3)

N-ras BP

OCT (2)

OCT (3)

ODC

p53 (3)

PAX-1

PAX-2 (2)

PAX-5 (2)

PAX-6 (2)

PBGD BP

PCF

PEBP-2 (2)

PEPCK PR GR

Pit-1 (2)

Pit-1 (3)

PO-B

PRDI-BFc

PRDII-BF1

PREB

PTF-1

PTF-1β

PU.1

PUR

RAR/RXR/CF1/

MB67

RFX-1/2/3 (3)

RORE

RVF

SAP-1b

Snail

SPERM-1

SRF (1)

SRF (2)

SSAP

Stat-1/3

Stat-5b

Surf-2 (2)

T3R

Tat

TFE-3L

TFEB

Thy-1 BP

tPA BP

Transferrin BP

TTF-1 (1)

TTF-1 (2)

TxREF/NF-lll

URE

WAP BP

XBP-1 (2)

XRE (2)

YB-1

Protein/DNA Array V

AP-1 (1)

AP-1 (2)

AP-2 (1)

AP-2 (2)

AR

Brn-3

CBF (1)

CDP

CEBP (1)

c-Myb (1)

CREB (1)

E2F-1 (1)

EGR-1 (1)

ER

Ets

Ets-1/PEA-3

FAST-1

GAS/ISRE

GATA

GR/PR

HNF-4 (1)

HSE

IRF-1

MEF-1

MEF-2 (1)

MRE

Myc-Max

NF-1 (1)

NFAT-1 (1)

NF-E1/YY1

NF-E2 (1)

NFκB (1)

OCT (1)

p53 (1)

PAX-5 (1)

Pbx-1

Pit-1 (1)

PPAR (1)

PRE

RAR/DR-5

RXR/DR-1

SIE

SMAD/SBE

SMAD-3/4

Sp-1 (1)

SRE

Stat-1

Stat-3

Stat-4

Stat-5

Stat-6

TFIID

TR

TR/DR-4

USF-1 (1)

VDR/DR-3

Protein/DNA Array I, TF-TF Interaction Array I, TF-Specific-TF Arrays I

Protein/DNA Array Content



NFkB p50-TF Interaction Array IMA5051 kitNFkB p50-TF Interaction Array IIMA5052 kitNFkB p50-TF Interaction Array IIIMA5053 kitNFkB p65-TF Interaction Array IMA5054 kitNFkB p65-TF Interaction Array IIMA5055 kitNFkB p65-TF Interaction Array IIIMA5056 kitPPARa-TF Interaction Array IMA5041 kitPPARa-TF Interaction Array IIMA5042 kitPPARa-TF Interaction Array IIIMA5043 kitPPARγ-TF Interaction Array IMA5044 kitPPARγ-TF Interaction Array IIMA5045 kitPPARγ-TF Interaction Array IIIMA5046 kit

�� A line of arrays for analysis of specific transcription factor complexes, focusing on interactions with NFkB and PPAR.

NFkB-TF Interaction Array NFkB-TF Interaction Array profiles interactions between NFkB and multiple TFs in a single hybrid-ization experiment and allows you to compare the interaction patterns under different conditions. Un-like traditional methods, this array-based technique generates a global view of interactions between NFkB and other proteins.

We currently offer three versions of the NFkB p50− and NFkB p65−TF Interaction Arrays, respectively, profiling a total of 244 different Transcription Factors (TFs). Each array is spotted with TF consensus-DNA binding sequences.

Here’s how they workThe TF-Specific-TF Interaction Arrays employ a pro-prietary, patent-pending technology that facilitates screening multiple interactions at once. The method is essentially the same as that used with the TF-TF Interaction Arrays, and the TF specific antibody is included for convenience.

References:1. Van Le, T. S., et al. (2004) Clin. Cancer Res. 10(4),

1384-1391.2. Hewetson, A., et al. (2003) J. Biol. Chem. 278(41),

40177-40185.

NFkB-TF and PPAR-TF Interaction Arrays

Nuclear extracts from untreated or TNFa-treated HeLa cells were incubated with the NFkB Probe mix. Immunoprecipitation with a NFkB-specific antibody was performed, and the protein-bound probes were then isolated and hybridized to the NFkB-TF Interaction Array membrane. Chemiluminescent images were collected using a FluorChem imager (Alpha Innotech), revealing changes in TF binding to NFkB upon TNFa stimulation.

Nuclear extracts from HeLa cells were incubated with the PPARa or PPARγ Probe Mix. Immunoprecipitation with the corre-sponding PPAR-specific antibody was performed, and the protein-bound probes were then isolated and hybridized to the PPARa or PPARγ-TF Interaction Array membrane. Chemiluminescent images were collected using a FluorChem imager (Alpha Innotech), revealing differences in PPARa- and PPARγ-interacting transcription factors in HeLa cells.

PPAR-TF Interaction ArrayPPAR-TF Interaction Array profiles interactions be-tween specific PPARs and multiple TFs. We currently offer three versions of the PPARa− and PPARγ−TF Interaction Arrays, respectively.

HeLa HeLa-TNFa

PPARa with HeLa extract PPARγ with HeLa extract



See chart on page 122 for full listing of Reporter Stable Cell Lines

�� Perform cell-based assays without transfection�� Screen compounds that inhibit or activate pathway components�� Monitor biochemical changes in NFkB, STAT, NFAT, CREB, AP-1, and SRF pathways

We offer NFkB, STAT-1, STAT-3, GR, NFAT, CREB, AP-1, SRF, TAD Kinase, and HIF Reporter Stable Cell Lines for high-throughput analysis of in vivo drug efficacy and specificity. These cells also provide a reproducible, ready-to-use system for performing cell-based assays. Among other applications, they can be used to evaluate uncharacterized growth factors, extracellular stimuli, and upstream events in the NFkB, STAT, NFAT, CREB, AP-1, SRF, TAD, and HIF signaling pathways. We currently offer NFkB Reporter Stable Cell Lines in five different cell lines: human A549, HeLa, 293, NIH3T3, and C2C12 cells. STAT-1 Reporter Stable Cell Lines are available in HeLa cells, NFAT in HeLa and K562, CREB in CHO and 293 cells, AP-1 in 293 cells, SRF in HeLa, and HIF in NIH3T3 cells.

All of the Reporter Stable Cell Lines maintain chromosomal integration of a luciferase reporter construct regulated by multiple copies of the transcription factor response element. The clonal cell lines are obtained by co-transfection of the luciferase reporter construct and pHyg followed by hygromycin selection.

NFkB p50 knockdownWe have established the NFkB p50 knockdown stable HeLa cells using a functionally validated shRNA expression vector. This cell line stably expresses siRNA specific for NFkB p50, and effectively knocks down the gene transactivation activity of NFkB. It is a powerful tool to identify novel NFkB target genes, and to analyze the interaction of other signaling proteins with NFkB. This stable cell line enables identification of downstream target genes of NFkB and allows the analysis of the cross-talk between NFkB and other signaling proteins.

NFkB Reporter Stable A549, HeLa, 293, NIH3T3 and C2C12 Cell LinesNuclear Factor kappa B (NFkB) is a member of the rel family of transcription factors. NFkB plays a key role in the regulation of inflammatory response, apoptosis, and tumorigenesis. NFkB is activated by a wide variety of different stimuli, including proinflammatory cytokines, oxidant free radicals, inhaled particles, ultraviolet radiation, and bacterial or viral products.

We have established NFkB reporter stable cells in a number of popular cell lines suitable for specific applications in the study of NFkB. They include 293 human embryonic kidney cells, HeLa human cervical epithelial cells, NIH3T3 mouse fibroblast cells, A549 human lung carcinoma cells, and C2C12 mouse myoblast cells. All these cell lines have been used extensively for the study of NFkB signaling due to their robust response. The 293 and HeLa cells are highly transfectable human cell lines whereas murine NIH3T3 cells are easy to maintain and transfect. In addition, A549 and C2C12 cells are useful models for studying the role of NFkB in lung carcinogenesis and muscle differentiation, respectively.

Reporter Stable Cell Lines

These cells have been treated with TNFa to induce lucifer-ase activity.

NFkB responsive luciferase reporter construct was trans-fected into the p50 knockdown stable cells or parental HeLa cells.


Luci

fera

se A

ctiv

ity

18,000

16,000

14,000

12,000

10,000

8,000

6,000

4,000

2,000

0

Parentalp50 KD

control

TNFα

Molecular Biology Products & Kits | Gene Regulation—Stable Cell Lines


Reporter Stable Cell Lines continued...

STAT-1 Reporter Stable HeLa Cell LineSTAT-1 is activated upon binding by IFN a, b, or γ; or IL-6 to its cognate receptor. Once STAT-1 is phosphorylated by JAKs, it forms a homodimer, or a heterodimer with STAT-2, and translocates to the nucleus. Once in the nucleus, the dimer binds to the GAS sequence (IFNγ activation site) in the promoter region of target genes and induces transcription.

These cells have been treated with IFNγ to induce luciferase activity.

AP-1 Reporter Stable 293 Cell LineThe activator protein 1 (AP-1) transcription factor is a dimeric complex that comprises members of the JUN, FOS, ATF, and MAF protein families with FOS and JUN being the most common of these proteins in mammalian cells. AP-1 activity is induced by growth factors, cytokines, and oncoproteins. Upon activation, AP-1 binds to the TPA Responsive ele-ment (TRE) and induces transcription of a variety of genes involved in multiple cellular functions such as proliferation, survival, differentiation and transforma-tion. The regulation of AP-1 activation is associated with the expression of genes important to cell differentiation programs.

CREB Reporter Stable CHO and 293 Cell LinesThe cyclic AMP response element (CRE)-binding protein (CREB) is one of the best-understood phos-phorylation-dependent transcription factors. Upon stimulation of cellular G-protein-coupled receptors and growth factor receptors, CREB is phosphory-lated by PKA, in a cAMP-dependent fashion. CREB can also be phosphorylated by a variety of other kinases in response to mitogen, calcium, and stress dependent signals. When phosphorylated on Ser133, CREB recruits transcriptional co- activators that induce transcription of a variety of intermediate early response genes. CREB is central to cellular processes such as growth factor-dependent cell proliferation and survival, and glucose homeostasis in learning and memory.

NFAT Reporter Stable HeLa and K562 Cell LinesNFAT transcription factors are activated by the calcium- and calmodulin-dependent serine phosphatase, calcineurin. Once dephosphorylated, NFAT translocates to the nucleus, where it acts synergistically with a wide variety of other transcription factors, such as AP-1, GATA, EGR, OCT, MEF-2, PPARγ, and HDACs. NFAT is subsequently phosphorylated and inactivated by a variety of kinases, such as GSK3, p38-MAPK, and PKA. The regulation of NFAT activation is associated with the expression of genes important to cell differentiation programs.

These cells have been tested with PMA to induce luciferase activity.

These cells have been tested with Forskolin to induce lucif-erase activity.

These cells have been treated with PMA and a calcium ionophore to induce luciferase activity.



Glucocorticoid Receptor Reporter 293 and HeLa Stable Cell LinesGlucocorticoids, a major subclass of steroid hormones, modulate a large number of metabolic, cardiovascular, immune, and behavioral functions. The intracellular effects of glucocorticoids are mediated by the glucocorticoid receptor (GR), a 94-kDa intracellular protein belonging to the phylogenetically conserved nuclear hormone receptor superfamily. In the absence of glucocorti-coid, the GR is maintained predominantly in the cytoplasm as part of an inactive multiprotein complex that consists of the receptor, two Hsp90 molecules, one molecule each of Hsp70 and Hsp56, and an immunophilin of the FK506- and rapamycin-binding class. When glucocorticoid binds to GR, the receptor undergoes a change in conformation, dissociates from regulatory heat shock proteins, and is hyperphosphorylated. The activated receptor rapidly translocates to the cell nucleus, where it is able to initiate transcriptional regulation.

In the nucleus, hormone-activated GR regulates transcription. During transcriptional regulation, the GR binds as a homodimer directly to short, palindromically arranged DNA sequences located in the promoter regions of glucocorticoid-responsive genes. Binding to these sequences, known as glucocorticoid response elements, leads to transcriptional induction or repression of target genes.

Induction of Luciferase Activity by Different Stimuli.

TAD-Kinase Reporter Stable Cell LineA previous system, the TAD in vivo Kinase Assay Kit, was developed to detect phosphorylation of the TAD of a specific TF. Now, to make this assay system more user-friendly, we have stably integrated the pGAL4-Luciferase vector into HeLa cells. Since only one vector, pGAL4-TAD, is needed for transfection into these cells, the sensitivity of this assay is dramatically increased. Upon stimulation, activated kinases will mediate modification of TADs leading to induction of the luciferase gene. Alternatively, kinases can be over-expressed via co-transfection with pGAL4-TAD and the activity of the kinase can be measured in vivo. Furthermore, the cells can be used in a mam-malian two-hybrid system for detecting protein-protein interaction.

These cells have been tested with Cobalt Chloride to induce luciferase activity.

SRF Reporter Stable HeLa Cell LineSerum response factor (SRF) is a widely expressed transcription factor involved in the expression of genes linked to cellular growth and muscle differen-tiation. Its target genes include cellular immediate early genes such as c-fos and structural cytoskeletal proteins, such as actins, myosins, tropomyosin, and vinculin. Serum-induced transcriptional activation of immediate early genes involves the formation of a ternary complex of SRF with a 62 kDa ternary complex factor (TCF) which binds to the serum response element (SRE). SRF is also involved in vascular smooth muscle differentiation by regulating the expression of several contractile and cytoskeletal genes. It is widely accepted that various diseases subvert normal smooth muscle cells to one of growth and excess matrix production.

These cells have been tested with Serum + PMA to induce luciferase activity.

HIF NIH3T3 Reporter Stable Cell LineHypoxia inducible factor (HIF) is a heterodimeric transcription factor composed of two basic helix-loop-helix proteins—HIF and HIF-1‚ (or Arnt). The HIFa/b dimer binds to hypoxia-response element and activates the transcription of many

genes that code for proteins that are involved in angiogenesis, glucose metabolism, cell proliferation/survival and invasion/metastasis, such as VEGF, FLT-1, EPO, and Leptin. Studies have shown the association of aberrant HIF activities with human diseases such as cancer and heart disease.

We have established HIF reporter stable cells in murine fibroblast NIH3T3 cells suitable for various applications in the research of HIF including the study of drug potency and the evaluation of novel inhibitor or activator proteins.

These cells have been tested with Cobalt Chloride to induce luciferase activity.

GRa CHO GFP Reporter Stable Cell LineThe CHO/ GFP-GRa cell line is designed for monitor-ing the activity of glucocorticoid receptor GRa in cell-based assays. The CHO/ GFP-GRa cell line was obtained by co-transfection of an expression vector for a fusion protein of TurboGFP (Evrogen PN FP511) and human GRa, as well as pHyg into Chinese ham-ster ovary (CHO) cells. Cells were then grown in the presence hygromycin. Single cells with strong green fluorescence were selected by flow cytometry, and allowed to expand. These cells constitutively express the GFP- GRa fusion protein.

NFkB CHO GFP Reporter Stable Cell LineThe CHO/ GFP-NFkBp65 cell line is designed for monitoring the activity of NFkB transcription factor in cell-based assays. The CHO/ GFP-NFkBp65 cell line was obtained by co-transfection of an expres-sion vector for a fusion protein of TurboGFP (Evrogen PN FP511) and human NFkBp65, as well as pHyg into Chinese hamster ovary (CHO) cells. Cells were then grown in the presence hygromycin. Single cells with strong green fluorescence were selected by flow cytometry, and allowed to expand. These cells constitutively express the GFP-NFkBp65 fusion protein.


a γ





Product Size Prod. no.AP-1 293 Reporter Stable Cell Line vial RC0006CHO/ GFP-GRa Stable Cell Line vial RC2002CHO/ GFP-NFkBp65 Stable Cell Line vial RC2001CREB 293 Reporter Stable Cell Line vial RC0007CREB CHO Reporter Stable Cell Line vial RC0008GR 293 Reporter Stable Cell Line vial RC0018GR HeLa Reporter Stable Cell Line vial RC0019HIF NIH3T3 Reporter Stable Cell Line vial RC0017NFAT HeLa Reporter Stable Cell Line vial RC0005NFAT K562 Reporter Stable Cell Line vial RC0009

Product Size Prod. no.NFkB 293 Reporter Stable Cell Line vial RC0014NFkB A549 Reporter Stable Cell Line vial RC0002NFkB C2C12 Reporter Stable Cell Line vial RC0016NFkB HeLa Reporter Stable Cell Line vial RC0013NFkB NIH3T3 Reporter Stable Cell Line vial RC0015NFkB p50 Knockdown HeLa Stable Cell Line vial RC5001SRF HeLa Reporter Stable Cell Line vial RC0012STAT-1 HeLa Reporter Stable Cell Line vial RC0004TAD-Kinase Reporter Stable Cell Line vial RC1001

Related products

Reporter stable cell lines can be stimulated with a variety of molecules. Some of these (cytokines, drugs, etc.) typically act at the cell surface, while others (siRNA, peptides) require transfection inside the cell.

By using the MPG amphipathic peptide rather than lipid mediated transfection, our DeliverX family of transfection reagents delivers siRNA or peptides to both cultured and primary cell lines with high efficiency and very low toxicity. See pages 98-100.

Reporter Stable Cell Lines



Molecular Biology Products & Kits | Next Generation Sequencing Library Prep



Generate high quality next generation sequencing (NGS) libraries suitable for sequencing on Illumina® platforms in less time with Prep2Seq DNA Library Prep Kit.

DNA library prep is critical to the precision and accuracy of NGS. The Prep2Seq DNA Library Prep Kit comes with all of the reagents necessary for end repair, A-tailing, and ligation, and can be completed in less than two hours.

Our kit is fast, efficient, and optimized. It efficiently converts 3’ and 5’ overhangs into 3’ A-overhang ends for ligation to T-tailed DNA adapters and reduces the potential for bias with our superior A-tailing and ligation steps. We offer significant improvement in the quality of sequence-ready libraries by eliminating chimeras and reducing side reaction fragments.

The Prep2Seq DNA Library Prep Kit combines end re-pair and A-tailing steps, decreasing the total number of steps to achieve quality NGS library preparation. Only one cleanup step is required to attain peak performance on the Illumina® sequencing platform and only 100 ng of input is needed.

Full kit offering�� Single kit that includes all reagents for sample prep in a slimmed down protocol.�� 24 bar code adapter sequences compatible with Illumina® technology are sold separately:

Prep2Seq Multiplex Oligo Adapters for Illumina - Index Adapter Set I–adapter sequences 1-12 - Index Adapter Set II–adapter sequences 13-27

Time savings with a more efficient method for NGS sample prep�� Improved A-tailing reaction with proprietary enzymes, cloning know-how, and optimized buffers.�� By combining the end repair and A-tailing steps, only one cleanup step is needed compared to the two or three steps in most other kits (saving up to two and a half hours in the lab).

The Prep2Seq DNA Library Prep Kit offers the highest adapter ligation efficiency available. By combining end repair and A-tailing, the Prep2Seq DNALibrary Prep Kit is able to reduce total NGS library prep time to 110 minutes and limit bead cleanup to a single step.

Rigorous comparison studies to competitor kits show that the Prep2Seq DNA Library Prep Kit generates impressive sequencing data.

Prep2Seq™ DNA Library Prep Kit for Illumina® platform

Table 1. Validated performance of the Prep2Seq DNA Library Prep Kit compared to competition.

USB® Prep2Seq™ Illumina® TruSeq™ NEBNext® Ultra™

Total Reads 170,507,429 171,139,925 176,073,965

Mapped reads to a single locus 142,223,440 143,567,898 147,688,157

% Mapped reads to a single locus 83.41% 83.89% 83.88%

Mapped reads to multiple loci 12,270,818 11,794,099 12,266,301

% Mapped reads to multiple loci 7.20% 6.89% 6.97%

Unmapped reads 16,013,171 15,777,928 16,119,507

% Unmapped reads 9.39% 9.22% 9.15%

The HiSeq® 2500 raw reads were aligned to the human genome hg19 release using Bowtie, allowing 2 mismatches for the entire read length. Mapped reads aligning to a single locus, mapped reads aligning to multiple loci, as well as unmapped reads from the 3 libraries are summarized in Table 1.

Table 2. GC content present in mapped and unmapped reads for the 3 library prep methods.


% GC: Mapped reads to a single locus 42.27% 40.01% 41.10%

% GC: Mapped reads to multiple loci 43.53% 41.65% 42.55%

% GC: Unmapped reads 40.90% 38.79% 40.64%

In order to determine if the end-repair and adaptor ligation may have contributed to some library bias we compared the overall GC content in mapped and unmapped reads for each of the 3 kits. As shown in Table 2, no substantial difference in GC content was introduced during the preparation methods of the 3 kits.

Time Comparison

from sheared DNA to size selection Illumina® TruSeq® Nano

140 min

NEBNext® Ultra™

65 min

USB Prep2Seq™

40 minHands-on Lab Time

Walk-Away Time

USB - 110 total min

NEB - 155 total min

Illumina - 210 total minIllumina® TruSeq® Nano

70 min

NEBNext® Ultra™

90 min

USB Prep2Seq™

70 min


Molecular Biology Products & Kits | Next Generation Sequencing Library Prep

Prep2Seq™ DNA Library Prep Kit for Illumina® platform continued...

79800 1 KT Set 1(Contains adapter sequences 1-12)

2 KT Set 2(Contains adapter sequences 13-27)

Prep2Seq Multiplex Oligo Adapters allow sample multiplexing in NGS reactions. These are HPLC purified adapter duplex sets that are bar coded and designed for multiplex sample preparation for Next Generation Sequencing on Illumina® platforms.

Set 1 components:12 multiplex adapters that correspond to Illumina®

adapters 1-12.20 reactions per adapter Each adapter vial contains 4.8 nmol, 80 μl.


adapters 13-16, 18-23, 25, and 27. 20 reactions per adapterEach adapter vial contains 4.8 nmol, 80 μl.

Prep2Seq Multiplex Oligo Adapters for Illumina® platform

The Prep2Seq DNA Library Prep Kit ensures efficiency of ligation to T-tailed adapters by optimizing the A-tailing step. This enzymology focus on A-tailing ensures that end bias is reduced, and the formation of blunt- and G-tailed ends is minimized. Most enzymes used for A-tailing are inefficient and result in a large amount of blunt-end DNA that can ligate as concatemers, forming chimeric false joints and compromising sequencing data. Effective A-tailing is crucial in preparing efficient DNA adapter libraries that are ready for sequencing.

Also available separately are barcoded adapter sets that enable multiplexing of up to 24 samples. Prep2Seq Multiplex Oligo adapters for Illumina® systems are available in kits of 1-12 and 13-27 duplex (79800 1 KT and 79800 2 KT).

Kit components:This kit is designed for input sheared genomic DNA amounts from 100 ng to 1 μg and consists of 3 tubes: Prep2Seq NGS Enzyme Mix, Prep2Seq NGS 2X Buffer, and Prep2Seq NGS High Concentration

Ligase. A streamlined workflow, straightforward pro-tocol, and high performing kit ensures quality library prep for your Illumina® sequencing runs.

Per 10 reactions: Prep2Seq NGS Enzyme Mix, 40 μl Prep2Seq NGS 2X Buffer, 250 μl Prep2Seq NGS High Concentration Ligase, 40 μl


WorkflowTime Comparison

from sheared DNA to size selection

min Ligation set up

NEB Next Ultra

USB Prep2Seq 5

min Ligation set up

30min Incubation

40min Incubation

60min Incubation 30min

Incubation55min

Clean up

5min

End repair set up

30min Incubation

35min

Clean up

5 30min Incubation

5

5 10min Incubation

90min Clean up

30min Clean up

5min End repair/ A-tail set up

min A-tail set up


210min TOTAL kit time



min Ligation set up

Illumina® TruSeq Nano®


78400 25 reactions78410 100 reactions

Description:The Ligate-IT Rapid Ligation Kit provides high-quality reagents for fast and easy ligation.

Quick and efficientLigation reactions are complete following room temperature incubation in just 5 minutes for cohesive-ends and 10 minutes for blunt-ends (Figs. 1 - 2). Transform competent cells immediately or store reactions at -20°C for later transformation.

ConvenientLigate-IT Rapid Ligation Kit includes T4 DNA Ligase, Nuclease-Free Water, and a specially formulated 5X Reaction Buffer which enhances the activity of T4 DNA Ligase, especially on blunt-end fragments. All of the components are in a ready-to-use format and do not require any additional reagents, such as ATP.

FlexibleThe kit can be used for a variety of ligation procedures such as conventional vector cloning, TA cloning, linker or adaptor ligation, and library construction.

Functional test:Cohesive- and blunt-end ligations of a 1200 bp insert into pUC19 yields ≥ 1 x 106 transformants per microgram of linearized, dephosphorylated vector.

Kit components:25 Reaction 100 Reaction

Pack Size Pack Size

Ligate-IT T4 DNA Ligase 25 µl 100 µl5X Ligate-IT Reaction Buffer 100 µl 400 µlWater, Nuclease-Free 1 ml 2 x 1 ml


Reference:1. Sambrook, J. and Russell, D. W. (2001) “Molecular


Related products:

FideliTaq DNA Polymerase71180 50 units 250 units 1,000 units 5 x 250 units 5,000 units

HotStart-IT FideliTaq DNA Polymerase71155 50 units 250 units 1,000 units 5,000 units




Ligate-IT Rapid Ligation Kit

Fig. 1. Rapid ligation of cohesive-end and blunt-end frag-ments. Lambda DNA was cut with Aat II (cohesive-end fragments) or EcoR V (blunt-end fragments) and purified. In each lane, 500 ng of cut lambda DNA were ligated with the Ligate-IT Rapid Ligation Kit for the indicated times (in minutes) and then loaded onto a 1% agarose gel. M are the DNA marker lanes. Results demonstrate fast and efficient ligation of both cohesive-end and blunt-end fragments in just 5 minutes.

Fig. 2. Ligate-IT Rapid Ligation Kit transformation efficiency of cohesive-end and blunt-end ligations. A modified pUC19 vector was cut to generate cohesive-ends (Aat II and EcoO109 I) and blunt-ends (Stu I). Both vectors were dephosphorylated prior to liga-tion. The lac repressor (lacI) from E. coli was cloned into the modified pUC19 vector with matching restriction sites for both cohesive-end and blunt-end ligations. Transformation efficiencies were determined by counting white colonies and are expressed in colony forming units per microgram of DNA used during transformation (cfu/µg). The uncut, supercoiled control vector was pUC19-lacI. Results indi-cate at least 106 transformants are obtained for both cohesive-end and blunt-end ligations when using competent cells that yield >108 cfu/µg.

Molecular Biology Products & Kits | Cloning


Molecular Biology Products & Kits | Mutagenesis

78480 20 reactions

The Change-IT Multiple Mutation Site Directed Mutagenesis Kit is designed to create single or multiple oligonucleotide-directed sequence changes in plasmids.

�� Based on highly efficient exponential amplification using non-overlapping primers; eliminates primer-dimer formation�� Create single or multiple mutations in plasmids�� Create mutations in plasmids as large as 15 Kb�� Create insertions, base changes, or deletions�� Create deletions as large as 300 bp�� Encode multiple base changes in a single primer�� Three step process: amplification, Dpn I digestion, and transformation�� Single day method

Mutations are created using a variation of ligation-during-amplification(1) in which the plasmid is PCR-amplified with phosphorylated primers and the product ends are ligated together to convert the annealed DNA strands into closed circular DNA molecules (Fig. 1). These closed circular DNA molecules then become template molecules in subsequent rounds of the PCR amplification. The desired mutations (insertions, deletions, or substitutions) are created by incorporating them into single or multiple primer sequences. FideliTaq™ Polymerase is used to ensure faithful duplication of the plasmid sequence and successful long-distance PCR. Thermostable DNA ligase is utilized for efficient creation of circular plasmid DNA during each round of PCR amplification. For convenience, the enzymes are included in the kit as Change-IT Enzyme. Change-IT Enzyme is a blend of FideliTaq DNA Polymerase and a thermostable ligase in 50% glycerol. Together these enzymes generate exponentially amplified, circular, mutated plasmid DNA. To enhance selection of the mutated plasmid from the parental plasmid, the PCR amplification reaction is digested with Dpn I prior to transforma-tion into competent cells.

ConvenienceMinimal hands-on time is required – set up an amplification reaction, a restriction enzyme digest, and a transformation. Kit includes optimized reac-tion buffer, phosphorylated common primers to b-lactamase, and enzymes.

High fidelity and large plasmidsFaithful amplification of plasmids is ensured by FideliTaq Polymerase, included in the Change-IT enzyme blend, and an optimized Change-IT Buffer. Mutate plasmids as large as 15 kb.

High efficiencyExponential amplification of the mutated plasmid provides for high mutagenesis efficiency, even with multiple mutations.

Table 1. Change-IT mutagenesis efficiency

Number of Mutations % Mutated

Single 90

Double 90

Triple 80

The efficiency for the creation of one, two, or three mutations in a single template were determined for pUC19 (Table 1). The single mutation blocked expression of b-galactosidase (Fig. 2). The double mutation blocked expression of b-galactosidase and created a Xho I site. The triple mutation created an Nco I site, a Xho I site, and blocked expression of b-galactosidase. The single and double mutations were created with the greatest efficiencies. The triple mutation was present in 80% of the plasmid product, a lower rate than the double mutation as expected for this technique.

Kit components:10X Change-IT BufferChange-IT EnzymeDpn IAmp FWD Primer: PO4/CCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAmp REV Primer: PO4/GTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGMutagenic Control Primer:PO4/ATGACCATGATTACGCCATAGCTTGCATGCCTGCAGG0.5 ng/μl pUC19Nuclease-Free Water

Shipping and storage:Shipped on dry ice. Store at -20ºC.

Reference1. Chen, Z. and Ruffner, D.E. (1998) Nucl. Acids Res.

26(4), 1126-1127.

Change-IT Multiple Mutation Site Directed Mutagenesis Kit

Fig. 1. Change-IT Mutagenesis Method. Phosphorylated oligo-nucleotides are annealed to template plasmid. One oligonucleotide bears the desired sequence changes while the other anneals to a common sequence, such as the b-lactamase ORF. FideliTaq DNA Polymerase extends the oligonucleotides and DNA ligase seals the nascent DNA strands creating a replicated plasmid bearing the desired mutation. This process is repeated in a PCR amplification for 15 to 30 cycles, generating exponentially amplified, mutated, double-stranded plasmid as product for subsequent transformation.

P

Anneal and Extend

Ligate Cycle

Dpn I Digestion

Transformation

P

P

P

Multiple mutations

Fig. 2. Creation of multiple mutations. Single, double, or triple mutation reactions were performed. The single mutation inactivated LacZa, the double mutation inactivated LacZa and added a Xho I site, and the triple mutation inactivated LacZa and added both an Nco I site and a Xho I site. Images of plates demonstrate that the LacZa inactivating mutation was present in all conditions (i.e. white mutated colonies pre-dominate). Colony PCR amplifications were then analyzed by restriction digests. The product from the single mutation condition lacked both Nco I and Xho I sites, as expected (single band visible on agarose gel). Restriction analysis of the double mutation condition revealed two bands in all colonies screened, indicating all carried the Xho I site insertion. Restriction analysis of the triple mutation condition revealed that 9 of 10 colonies incorporated all three site-directed mutations.


Molecular Biology Products & Kits | Labeling

78430 30 reactions

The Sequenase Random Primer Labeling Kit offers a rapid and convenient method to generate DNA probes with high specific activity, >1.5 x 109 dpm/μg. The kit features Sequenase Version 2.0 DNA Polym-erase and random nonamers (9-mers) in a protocol that has been optimized to produce radiolabeled probes in 5 minutes. The labeled probes may be used for screening of gene libraries(1), Southern and North-ern blots(2,3), as well as in situ hybridizations.

�� Rapid, 5-minute reaction�� DNA probes labeled to high specific activity, >1.5 x 109 dpm/μg�� Easy-to-use 10X Nucleotide Mixes�� Flexible labeling with either radiolabeled dATP or dCTP�� Optimized labeling conditions using Sequenase Version 2.0 DNA Polymerase and random nonamers

Random primer labeling methodRandom primed labeling of DNA was originally developed by Feinberg and Vogelstein and used Klenow DNA Polymerase as the enzyme for polym-erization(4,5). The method relies on annealing random DNA primers along the length of a target DNA so that the primed DNA can be used as a substrate for the polymerase. The DNA is radiolabeled by substituting one of the nucleotides of the reaction with a radioactive one ([32P], [33P], or [35S]), and the resulting radiolabeled DNA can be used as probes in various hybridization techniques such as Southern and Northern blots and in situ hybridizations. This original protocol has been enhanced by labeling probes with Sequenase DNA Polymerase instead of Klenow(6). Sequenase DNA Polymerase is the enzyme of choice because of its superior activity in primer extension reactions and its lack of exonuclease activ-ity that tends to degrade radiolabeled probes. Also, the kit includes random nonamers (9-mers) instead of hexamers (6-mers) for the labeling reaction to provide more efficient priming of the target DNA(7).

Applications:�� Southern blots�� Northern blots�� Solution hybridization�� In situ hybridization

High specific activity probesThe Sequenase Random Primer Labeling Kit suc-cessfully labels varying amounts of DNA up to approximately 2 μg, but 25 ng is recommended for maximal probe specific activity. Figure 1 shows a time course of the labeling reaction using 25 ng of Lambda DNA that was monitored by the incorpora-tion of [33P]-dATP. The standard protocol routinely gives a specific activity of >1.5 x 109 dpm/µg within five minutes, corresponding to an incorporation of up to 70% of the radioactive nucleotide.

Consistent probe sizeThe Sequenase Random Primer Labeling Kit gener-ates a consistent range of probe sizes on a variety of DNA templates, including lambda DNA, pUC18 plasmid DNA, as well as DNA fragments gener-ated by PCR (Fig. 2). The labeling protocol routinely produces DNA probes ranging in size from 100-850 nucleotides in length.

Easy to useThe Sequenase Random Primer Labeling Kit provides two different 10X Nucleotide Mixes (-dATP and -dCTP) for flexible labeling. Using these buffers, DNA probes can be generated equally well using either radioactively labeled dATP or dCTP.

Kit components: 10X Primer Mix10X Nucleotide Mix, minus dATP10X Nucleotide Mix, minus dCTPSequenase Version 2.0 DNA PolymeraseWater, Nuclease-FreeStop Buffer (0.5 M EDTA)Control DNA (lambda DNA)

Shipping and storage:Shipped on dry ice. Store at -20ºC.

References: 1. Grunstein, M. and Hogness, D. S. (1975) Proc.

Natl. Acad. Sci. USA 72, 3961-3965.2. Southern, E. M. (1975) J. Mol. Biol. 98, 503-517.3. Smith, G. E. and Summers, M. D. (1980) Anal.

Biochem. 109, 123-129.4. Feinberg, A. P. and Vogelstein, B. (1983) Anal.

Biochem. 132, 6-13.5. Feinberg, A. P. and Vogelstein, B. (1984) Anal.

Biochem. 137, 266-267.6. U.S. Pat. No. 4,795,699.7. Stangegaard, M., Dufva, I. H., and Dufva, M.

(2006) BioTechniques 40, 649-657.

Sequenase Random Primer Labeling Kit

Fig. 2. Size comparison of DNA probes generated from lambda DNA (Lane 1), linearized pUC18 (Lane 2), and a PCR fragment, 1.6 kb (Lane 3).

Fig. 1. Specific activity of DNA probes generated from lamb-da DNA through time.

Minutes

Spec

ific

Activ

ity(x

10

9dp

m/

g)

1510500.0

0.5

1.0

1.5

2.0

2.5

850 —

500 —400 —300 —200 —

100 —

DNA probesMW (nt)

1 2 3

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Denhardt’s Solution, 50X70468 50 ml

SSC, 20X Solution19629 500 ml 1 L 5 L

SSPE, 20X Solution75890 1 L 5 L




DNA, Calf ThymusULTRAPURE

Source: Calf ThymusDescription: Highly polymerized DNA; Characterized

as a high molecular weight DNA prepared by ethanol precipitation. This DNA is an excellent substrate for DNase.

Product specifications:Assay: Hyperchromicity ≥ 27%A260/A280: ≥ 1.8

Storage: 4°C, desiccatedCAS #9007-49-2

14375 1 gm 5 gm

DNA, E. coli

Source: E. coliDescription: E. coli genomic DNA is purified from

E. coli type B cells, ATCC 11303 strain. It has a single chromosome and the genome is 4,600,000 bp long. This genomic DNA is fragmented to some degree during purification yet it is charac-terized as a high molecular weight DNA.

Product specifications:Form: Dried powderHyperchromicity: ≥ 27%A260/A280: ≥ 1.8

Storage: 4°CCAS #9007-49-2

14380 10 mg

DNA, Fish Sperm

Source: Fish SpermProduct specifications:Form: Off-white to yellowish/brownish powderMoisture: ≤ 7%Nitrogen: ≥ 14.5%Phosphorous: ≥ 9.0%Protein: NegativeCAS #100403-24-5

14405 100 gm 1 kg

DNA, Fish Sperm, Activated

Form: Solution in 10 mM Tris-HCI, pH 7.5, 1 mM EDTA.

Activity Analysis: This DNA gives the optimum activ-ity in standard assay for Sequenase Version 2.0 DNA Polymerase.

Storage: -20°C

70076 500 µg 2 mg

DNA, Fish Sperm, Sodium Salt

Source: Fish SpermProduct specifications:Form: Off-white to light tan powderPhosphorous: ≤ 9.0%Protein: ≤ 5.0%Loss on Drying: ≤ 18.0%CAS #68938-01-2

14377 100 gm 1 kg

DNA-Cellulose, Double-Stranded(dsDNA-Cellulose)

Source: Calf ThymusDescription: Calf Thymus DNA (PN 14375) and ash

free, acid washed, chromatographic grade cellu-lose and crosslinked by ultraviolet irradiation.

Product specifications:Form: Dried powder (may contain traces of NaCl,

Tris-HCl and EDTA).Use: Column chromatography media for the iso-

lation of DNA binding proteins from various sources.

Note: Store as dry powder after washing with H2O and ethanol.

Note: One gram of DNA Cellulose will swell to 3-4 ml gel when fully hydrated.

CAS #9004-34-6

14394 5 gm 10 gm

Molecular Biology Products & Kits | DNA

We have been providing custom enzymes and bulk biochemicals for more than 40 years, helping academic institutions, diagnostic, biotech, and pharmaceutical companies succeed. We combine the resources of a large organization with the personalized service of a dedicated team. Turn to us for:

� Custom reagent manufacturing, dispensing, labeling, and kitting

� 3,000 catalog enzymes and biochemicals that can be tailored to meet your specific needs

� An ISO 13485 certified quality system well-suited to meet diagnostic requirements

� In-house experts in custom and OEM reagents to guide you through the manufacturing process


DNA-Cellulose, Single-Stranded(ssDNA-Cellulose)

Source: Calf ThymusDescription: Calf Thymus DNA (PN 14375) and ash

free, acid washed, chromatographic grade cellu-lose and crosslinked by ultraviolet irradiation.

Product specifications:Form: Dried powder (may contain traces of NaCl,

Tris-HCl and EDTA). One gram of powder swells to approx. 3 ml of final hydrated media.

Note: Store as dry powder after washing with H2O and ethanol.

Use column chromatography media for the isolation of DNA binding proteins from various sources.

Note: One gram of DNA Cellulose will swell to 3-4 ml gel when fully hydrated.

CAS #9004-34-6

14395 5 gm 10 gm

M13mp18, Single-Stranded, 0.2 µg/µl

Source: E. coli strain JM101 infected with bacterio-phage M13mp18

Storage Buffer: Supplied in 10 mM Tris-HCl (pH 8.0), 1 mM EDTA.

Concentration: 0.2 µg/µl

70704 50 µl 50 µg

M13mp18 DNA, Single-Stranded, 1.0 µg/µl

Source: E. coli strain JM101 infected with bacterio-phage M13mp18


Concentration: 1.0 µg/µl

71706 50 µg 200 µg

Oligo(dT)12-18 Primer, MW ≅ 4500

MB GradeForm: 25 µM ± 2.5 in dH2O.2.5 nmol per tube.Storage: -20°C.

77405 100 µl (2.5 nmol)

Oligo p(dT)12-18 Sodium Salt

Form: Lyophilized powderDescription: This oligodeoxythymidylate mixture is

synthesized using O-cyanoethyl phosphoramidite chemistry. After the removal of the protecting groups, the product is converted to the sodium salt and the excess salt removed by gel filtration.

Unit Definition: One A260 unit is that quantity of material which, when dissolved in 1.0 ml of water and read in a spectrophotometer at 260 nm in a cuvette with a 1.0 cm light path, gives an absor-bance of 1.0. One A260 unit is approximately equal to 30-35 µg of DNA.

Storage: -20°C

19817 5 units 25 units 100 units

pUC19 DNA, 0.2 µg/µl


Concentration: 0.2 µg/µlFunctional Test: Tested in DNA sequencing

76632 10 µg 50 µg 200 µg

pUC19 DNA, 1.0 µg/µl


Concentration: 1.0 µg/µlFunctional Test: Tested in DNA sequencing

76634 50 µg 200 µg

Custom sizes, concentrations and pack sizes available upon request

Molecular Biology Products & Kits | DNA


Molecular Biology Products & Kits | DNA Markers

76710 250 µl

6X DNA Loading Buffer (OXG)76715 1 ml 5 ml

Description:PCR Markers are a mixture of 8 DNA bands ranging in size from 50 to 2000 bp in ready-to-load form supplied in gel loading buffer. The recommended amount for loading (5 µl) produces bands of high resolution for easy size estimation of PCR products.

6X DNA Loading Buffer (OXG) with orange G and xylene cyanol FF is included for convenience to pre-pare PCR samples for agarose gel electrophoresis. The tracking dyes do not interfere with UV illumina-tion of ethidium bromide stained gels.

Fragments: 2000 bp 500 bp 1500 bp 300 bp 1000 bp 150 bp 750 bp 50 bp

To use: Add 5 µl PCR Marker per gel lane.

6X DNA Loading Buffer (OXG) (PN 76715, 1 ml included):6X Loading Buffer consists of orange G, xylene cyanol FF, and glycerol. Shipped ambient.

Functional test:Meets or exceeds criteria for agarose gel electro-phoresis.

Shipping and storage:Shipped on ice. Store at 4°C.

PCR Markers, 50 - 2000 bp

– 2000 – 1500

– 1000 – 750

– 500

– 300

– 150

– 50

bp

1.5% TAE agarose gel

76712 500 µl

6X DNA Loading Buffer (OXG)76715 1 ml 5 ml

Description:The 100 bp DNA Ladder is a set of 10 easy-to-re-member band sizes for use in qualitative agarose gel analysis. The ladder is supplied in ready-to-load form in gel loading buffer. Simply add the recommended amount (5 µl) for bright band intensities between 100 and 1000 base pairs.

6X DNA Loading Buffer (OXG) with orange G and xylene cyanol FF is included for convenience to prepare DNA samples for agarose gel electrophore-sis. The tracking dyes do not interfere with UV illumination of ethidium bromide stained gels.

Fragments: 1000 bp 500 bp 900 bp 400 bp 800 bp 300 bp 700 bp 200 bp 600 bp 100 bp

To use: Add 5 µl of ladder mix per gel lane.

6X DNA Loading Buffer (OXG) (PN 76715, 1 ml included):6X Loading Buffer consists of orange G, xylene cyanol FF, and glycerol. Shipped ambient.



DNA Ladder, 100 bp


– 1000 – 900 – 800 – 700 – 600 – 500

– 400

– 300

– 200

– 100

bp


Molecular Biology Products & Kits | DNA Markers

76714 500 µl

6X DNA Loading Buffer (BXF)76720 1 ml 5 ml

Description:The 1 kb Plus DNA Ladder is a set of 14 easy-to-remember band sizes for use in qualitative agarose gel analysis ranging in size from 12 kb down to 100 bp. Two high intensity reference bands of 4 kb and 500 bp are included. The ladder is supplied in ready-to-load form in gel loading buffer. Simply add the recommended amount (5 µl) for highly resolved bands.

6X DNA Loading Buffer (BXF) with bromophenol blue and xylene cyanol FF is included for conve-nience to prepare DNA samples for agarose gel electrophoresis.

Fragments: 12 kb 1.5 kb 10 kb 1 kb 8 kb 500 bp 6 kb 400 bp 4 kb 300 bp 3 kb 200 bp 2 kb 100 bp

To use: Add 5 µl ladder mix per gel lane.

6X DNA Loading Buffer (BXF) (PN 76720, 1 ml included):6X Loading Buffer consists of bromophenol blue, xylene cyanol FF, and Ficoll®.



DNA Ladder, 1 kb Plus


– 12,000 – 10,000 – 8000 – 6000

– 4000 – 3000

– 2000

– 1500

– 1000

– 500 – 400 – 300 – 200 – 100

bp

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Molecular Biology Products & Kits | Oligonucleotide Marker

76410 100 µl

Description:The Low Molecular Weight Marker is a set of 14 bands between 10 - 100 nucleotides, designed for size determination of single-stranded oligonucleotide fragments.

The marker is suitable for use in either polyacryla-mide or agarose gel electrophoresis. The recom-mended concentration for agarose is 2.5% and for polyacrylamide is 12 - 15%. This marker should be diluted with TE Buffer and the supplied Gel Loading Buffer to the desired loading concentration.

The Low Molecular Weight Marker can be used to generate a 5’-end radiolabeled marker for size comparison of small, single-stranded oligonucleotide fragments such as miRNA, siRNA, tRNA, snRNA, snoRNA, or ssDNA. The Low Molecular Weight Marker is also useful when the molecule of interest is less than 100 nucleotides in length. This marker is not designed for precise quantification of nucleic acid mass and should only be used as a size marker.

2X Formamide Loading Buffer and 6X Glycerol Load-ing Buffer are included for convenience to prepare both the marker and samples for gel electrophoresis.

Fragment sizes: 100 nt 40 nt 90 nt 35 nt 80 nt 30 nt 70 nt 25 nt 60 nt 20 nt 50 nt 15 nt 45 nt 10 nt

Functional test: Meets or exceeds criteria for 5'-end labeling with isotope and polyacrylamide gel electrophoresis.

Components:Low Molecular Weight Marker, 100 µl (50 ng/µl each

fragment)2X Formamide Loading Buffer6X Glycerol Loading Buffer

2X Formamide Loading Buffer (1 ml included):Consists of bromophenol blue, xylene cyanol FF, EDTA and formamide.

6X Glycerol Loading Buffer (1 ml included): Consists of orange G, xylene cyanol FF and glycerol.

Shipping and storage:Shipped on ice. Store at 4°C or at -20°C for long term storage.

To use:1. Mix 2 µl (100 ng each fragment) of Marker with

2 µl of 2X Formamide Loading Buffer, heat at 95°C for 3 minutes, quick cool on ice, and load the entire volume per lane on polyacrylamide gel.

2. Mix 5 µl (250 ng each fragment) of Marker with 1 µl of 6X Glycerol Loading Buffer and load the entire volume per lane on agarose gel.

3. Visualize with dyes for detecting single-stranded oligonucleotide fragments.

4. For 5'-end labeling reaction, use 1 µl (50 ng each fragment) of Marker with OptiKinase™ and [γ-32P]-ATP or [γ-33P]-ATP.

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5 L

TE Buffer, 1X Solution75893 100 ml 10 x 1 ml

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TEMED76320 100 ml 100 gm

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Low Molecular Weight Marker, 10 – 100 nt

Fig. 1. Low Molecular Weight Marker on 12-15% UREA-polyacrylamide gel. A. 50 ng of SYBR® Gold stained Marker. B. 0.5 ng of 5' end labeled Marker with [γ-33P]-ATP and OptiKinase. C. 0.05 ng of 5' end labeled Marker with [γ-32P]-ATP and OptiKinase.

100908070

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5045

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100908070

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10090807060

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A B Cnt nt nt


Molecular Biology Products & Kits | Protein Analysis

Human Angiogenesis Antibody ArrayMA6310 kitMouse Angiogenesis Antibody ArrayMA6320 kit

�� Simultaneously profile expression of multiple proteins associated with angiogenesis�� Analyze the effects of different stimuli on the induction or inhibition of angiogenesis

With Angiogenesis Antibody Arrays, you can detect multiple angiogenesis-specific cytokines in a single hybridization experiment. These arrays are extremely sensitive, detecting proteins at concentrations in the pg/ml range.

Angiogenesis, the generation of blood vessels from endothelial cells, is normally controlled by pro-angio-genic “activator” molecules and anti-angiogenic “inhibitor” molecules. In some cancers, however, the careful balance of these molecules is upset, as tumor cells promote vascularization to obtain nutrients and oxygen necessary for growth. Tumor growth is sustained by abnormal levels of these angiogenesis-related proteins; the affected proteins vary from one type of cancer to another. To gain a clear picture of the angiogenesis-stimulating pathways used by cancer cells, it is necessary to investigate the levels of these cytokines in tumors. With the Angiogenesis Antibody Arrays, you can simultaneously profile 19 angiogen-esis activators and inhibitors at the protein level.

We currently offer Angiogenesis Antibody Arrays for both human and mouse. These arrays can be used with a variety of biological sources, including cell extracts, tissue lysates, conditioned media, patient sera, and plasma.

Kit components:Four array membranesBiotinylated angiogenesis antibodiesBlocking, hybridization, and wash solutionsHRP detection reagents

Angiogenesis Antibody Array content Here’s how it works

Angiogenesis Antibody Arrays

Human Angiogenesis Array

Angiogenin

G-CSF

HGF

Leptin

VEGF

IL-1a

IL-1b

IL-6

IL-8

PIGF

FGFa

FGFb

TNFa

TGFa

IFNγ

IL-12

IP-10

TIMP-1

TIMP-2

Mouse Angiogenesis Array

EGF

FGFa

FGFb

G-CSF

IL-1a

IL-1b

IL-4

IL-6

Leptin

VEGF

TNFa

TGFa

TGFb

IFNγ

IL-12

IP-10

TIMP-1

TIMP-2

The Angiogenesis Antibody Array profiles expression of angiogenesis-specific cytokines. Tissue extracts from normal (Panel A) and cancerous (Panel B) human colon were incubated with the Angiogenesis Antibody Array membrane. Chemiluminescent images were collected using a FluorChem imager (Alpha Innotech), revealing increases in the expression of certain cytokines in the can-cerous tissue extract (boxes).

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Looking to profile cytokines from a larger number of samples? Call our Technical Support team and ask about the Procarta Cytokine Plex Profiling Kits. Both human and mouse panels are available.


AFP ELISA Kit BC1009 kitCA125 ELISA Kit BC1013 kitCA15-3 ELISA Kit BC1015 kitCA19-9 ELISA Kit BC1017 kit

Quickly quantify cancer antigen concentra-tions in human serumCancer Antigen ELISA Kits allow rapid screening of up to 96 samples at once for quantitative determina-tion of tumor marker concentration. This ELISA technology allows you to quickly and efficiently screen multiple human serum samples for various antigens shown to be present at elevated levels in different cancers. The four assays we offer are for alpha-fetoprotein (AFP) and cancer antigens CA125, CA15-3, and CA19-9.

Note: The Cancer Antigen ELISA Kits are intended for research use only.

Kit components:One 96-well microtiter plate coated with anti-cancer

antigenProtein standardsComplete reaction and detection reagents

AFPElevation of serum AFP to abnormally high levels occurs in several malignant diseases (including nonseminomatous testicular cancer and primary hepatocellular carcinoma) and some benign ones (including hepatitis and cirrhosis).

CA125Cancer Antigen 125 (CA125) is a surface antigen associated with epithelial ovarian cancer, and to date CA125 is the most sensitive marker for residual epi-thelial ovarian cancer. CA125 may also be elevated in patients with lung, cervical, fallopian tube, and uterine cancers, as well as endometriosis.

CA15-3Cancer antigen 15-3 (CA15-3) is useful for monitor-ing breast cancer patients post-operatively for recurrence, particularly with metastatic disease. CA15-3 levels are also increased in colon, lung, and hepatic tumors.

CA19-9Serum CA19-9 level is frequently elevated in subjects with certain gastrointestinal malignancies, such as pancreatic, colorectal, gastric, and hepatic carcino-mas. A consistently rising serum CA19-9 value may be associated with progressive malignant disease and poor therapeutic response. A declining CA19-9 value may be indicative of a favorable prognosis and good response to treatment.

Cancer Antigen ELISA Kits

AFP

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

0 50 100 150 200 250 300

Concentration ng/mlA

450

A45

0

CA125

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

0 100 200 300 400

Units/ml

A45

0

CA15-3

Units/ml

0.0

0.5

1.0

1.5

2.0

2.5

3.0

0 50 100 150 200 250

CA19-9

A45

00.0

0.5

1.0

1.5

2.0

2.5

3.0

0 100 200 300 400 500 600

Units/ml

Human Cytokine Antibody Array 1.0MA6110 2 membranesHuman Cytokine Antibody Array 1.0MA6120 4 membranesHuman Cytokine Antibody Array 1.0MA6130 6 membranesHuman Cytokine Antibody Array 1.0MA6140 8 membranesHuman Cytokine Antibody Array 3.0MA6150 4 membranesHuman Cytokine Antibody Array 3.0MA6160 8 membranesMouse Cytokine Antibody Array 1.0MA6410 4 membranesMouse Cytokine Antibody Array 1.0MA6412 8 membranes

�� Simultaneously profile the expression of multiple cytokine proteins.�� Analyze regulation of cytokine expression with different cellular stimuli.

With the Cytokine Antibody Arrays, you can simul-taneously detect multiple cytokines from a variety of sources, including serum, conditioned media, plasma, and cell/tissue lysates. While cDNA arrays give information about levels of gene expression, the Cytokine Antibody Arrays detect the levels of active cytokine protein.

The Cytokine Antibody Arrays have several advan-tages over traditional methods such as ELISA. The sensitive high-throughput assay is a key benefit—these arrays can detect soluble cytokine proteins at concentrations in the pg/ml range. There is also less variability with these arrays compared to ELISAs; variability between the same spots on two of the same type of membrane is typically less than 10%, while the variation between ELISA plates is about 20%. Most importantly, the Cytokine Antibody Ar-rays facilitate the simultaneous detection of multiple cytokines in one assay.

We currently offer three of these arrays—two for human and one for mouse. The Human Cytokine Antibody Array comes in two formats: Version I.0 includes 18 cytokines and Version 3.0 includes 36 cytokines. The Mouse Cytokine Antibody Array 1.0 includes 18 cytokines.

The Cytokine Antibody Arrays can be purchased with different numbers of membranes.

Kit components:Array membranesBiotinylated anti-cytokine antibodiesStreptavidin-HRP conjugateBlocking and wash solutionsHRP detection reagents

Cytokine Antibody Arrays



The Human Cytokine Antibody Array 3.0 detects differences among active cytokines from A431 cells exposed to different conditions. Membranes were incubated with conditioned media from untreated A431 cells (Panel A) and A431 cells treated with EGF (Panel B) or PMA (Panel C). Images were collected using a FluorChem imager (Alpha Innotech). Differences in protein expression were observed among the untreated and treated samples, as indicated in boxes.

B. Unstimulated A431 and EGF C. Stimulated A431 and PMAA. Unstimulated A431

Human Cytokine Array 1.0

EotaxinIL-1aIL-10GM-CSFIL-1bIL-12IP-10IL-3IL-17TNFaIL-4MIP1aRantesIL-6MIP1bLeptinIL-8MIP-5

Mouse Cytokine Array 1.0

G-CSFM-CSFGM-CSFMIGMIP-1aIFNγ

TNFaIP-10RantesVEGFIL-1aIL-2IL-4IL-5IL-6IL-10IL-12IL-13

Human Cytokine Array 3.0

Apol/FasCTLAEotaxinGM-CSFEGFIP-10LeptinMIP1aMIP1bMIP4MIP-5MMP3RantesTGFbIFNγ

TNFaTNFRITNFRIIICAM-1VCAM-1VEGFIL-1aIL-1bIL-1RaIL-2IL-3IL-4IL-5IL-6IL-6RIL-7IL-8IL-10IL-12 (p40)IL-15IL-17


Cytokine Antibody Arrays continued...

Cytokine Antibody Content



76740 500 µl

Description:The 10 - 225 kDa Protein Markers are recombinant proteins designed for convenient and accurate sizing of protein samples. Unlike conventional protein markers, the recombinant marker proteins do not contain oligosaccharides which often cause poor resolution and inaccurate size estimation of protein bands.

An added benefit is the ability to estimate the con-centration of protein samples using the known mass for each protein band. Each vial (500 µl) contains 500 µg of protein - 50 µg per band except for 100 µg of the 50 kDa high intensity reference band.

The markers are in ready-to-load form in gel loading buffer. Simply add 5 µl of the marker to a standard SDS-polyacrylamide gel for visualization with Coomassie blue.

As an added convenience, 2X SDS Sample Buffer is included for preparing protein samples for gel electrophoresis.

Protein bands (MW): 225 kDa 35 kDa 150 kDa 25 kDa 100 kDa 15 kDa 75 kDa 10 kDa 50 kDa

The 50 kDa band is a high intensity reference band.

Storage buffer:1 µg/µl total marker protein in 125 mM Tris-HCl, pH 6.8, 2% SDS, 10% glycerol, 200 mM b-ME, 0.007% bromophenol blue.

To use: Add 5 µl of marker per gel lane on standard SDS-polyacrylamide gels for staining with Coomassie blue. For silver staining or S-Taq analysis, use 5 µl of a 1:25 dilution in 1X Sample Buffer.

2X SDS Sample Buffer (1 ml included)250 mM Tris-HCl, pH 6.8, 6% SDS, 300 mM DTT, 30% glycerol, and 0.02% bromophenol blue.

Functional test:Meets or exceeds criteria for SDS-PAGE.


Related products:

Acrylamide75820 100 gm 500 gm 1 kg

Ammonium Persulfate76322 10 gm 100 gm 1 kg

N,N'-Methylene-bis-Acrylamide75821 25 gm 50 gm 100 gm 1 kg

RapidGel, 40% Liquid Acrylamide Stock Solution75848 500 ml

Sodium Dodecyl Sulfate, 10% Solution77504 100 ml 500 ml 1 L

TBS with Tween (TBST), 20X Solution77500 1 L 5 L

TEMED76320 100 gm 100 ml 500 gm

Tris-Glycine (TG) Buffer, 10X Solution75894 1 L 5 L

Tris-Glycine-SDS (TGS) Buffer, 10X Solution75895 1 L 5 L

Protein Markers, 10 - 225 kDa

Stained with Coomassie Blue 8-16% Tris-Glycine gel.

kDa – 225

– 150

– 100

– 75

– 50*

– 35

– 25

– 15 – 10

Molecular Biology Products & Kits | Protein Markers


76230Y 100 mg76230Z 500 mg76230 1 gm (2 x 500 mg)Customs by request

Source:Tritirachium album

Description:Proteinase K is a highly active and stable endopep-tidase purified from the fungus T. album. It cleaves peptide bonds mainly following the carboxyl group of N-substituted hydrophobic aliphatic and aromatic amino acids(1) and is classified as a serine protease. Proteinase K is useful in purifying high molecular weight nucleic acids from cells and tissues (i.e. genomic DNA isolation from mouse tails) and in revealing protein structure and function.

Applications:�� Used in the isolation of native, high molecular weight DNA and RNA.�� Used as a general protease to inactivate DNases, RNases, and to degrade proteins.�� Specifically modifies cell surface proteins and glycoproteins for analysis of membrane structures for protein localization(5,6).�� Produces characteristic protein fragments used in enzyme/protein structure and function studies(7).

Form:Lyophilized powder

Properties:Molecular Weight: 28.9 kDaOptimum pH: 4.0-12.5(1)

Optimum Temperature: 65°C(1)

Working Concentration: 0.05 - 1 mg/mlActivators: Stimulated by 0.2 - 1% SDS or 1 - 4 M

urea(8) and 1 - 5 mM Ca2+

Inactivated by PMSF and diisopropyl phosphoro-fluoridate (DPF). Partially inactivated by EGTA. Not inactivated by EDTA, thiol reagents, or trypsin/chymotrypsin inhibitors(1)

Note: Proteinase K has two binding sites for Ca2+ which act as a stabilizing factor of the enzyme. When Ca2+ is removed from the enzyme approxi-mately 80% of the catalytic activity is lost because of the structural change(2). However, residual Proteinase K activity is usually sufficient to degrade proteins commonly present during DNA/RNA isolation.

Purity:Tested for contaminating ribonucleases and deoxy-ribonucleases.

Unit definition:One unit is the amount of enzyme that liberates folin positive amino acids and peptides corresponding to

1 µmol tyrosine in 1 minute at 37°C using hemoglo-bin as substrate.

Concentration:20 - 50 units/mg

Shipping and storage:Shipped on cold pack. Store dry at 4°C.

References:1. Ebeling, W., Hennrich, N., Klockow, M., Metz, H.,

Orth, H. D. and Lang, H. (1974) Eur. J. Biochem. 47, 91-97.

2. Gross-Bellard, M., Oudet, P. and Chambon, P. (1973) Eur. J. Biochem. 36, 32-38.

3. Hansen, J. N. (1974) Prep. Biochem. 4, 473-488.4. Kasche, V., Zollner, R., Amneus, H. and Naslund, L.

(1981) Prep. Biochem. 11, 233-250.5. Roelcke, D. and Ulenbruck, G. (1969) Z. Med.

Mikrobiot. und Immunol. 155, 156-170.6. Brdiczka, D. and Krebs, W. (1973) Biochem.

Biophys. Acta 297, 203-212.7. Williamson, J., Greene, J., Cherif, S. and Milner-

White, E. J. (1977) Biochem. J. 167, 731-737.8. Hilz, H., Wiegers, U. and Adamietz, P. (1975) Eur. J.

Biochem. 56, 103-108.9. Crowe, J. S., Cooper, H., Smith, M., Sims, M.,

Parker, D. and Gewert, D. (1991) Nucl. Acids. Res. 19, 184.

Proteinase K

76225 100 mg

Source:Tritirachium album

Description:Proteinase K is a highly active and stable endopep-tidase purified from the fungus T. album. It cleaves peptide bonds mainly following the carboxyl group of N-substituted hydrophobic aliphatic and aromatic amino acids(1) and is classified as a serine protease. Proteinase K is useful in purifying high molecular weight nucleic acids from cells and tissues and in revealing protein structure and function.

Applications:�� Isolation of DNA and RNA to inactivate DNases, RNases and to degrade other proteins during cell digestion.�� Useful in the specific modification of cell surface proteins and glycoproteins for analysis of membrane structures(4,5).�� Useful in obtaining characteristic protein fragments which help reveal the structure and function of enzymes and other proteins(6).

Properties:Molecular Weight: 28,900 DapH Range: 4.0 -12.5(1) with optimum activity at

pH 8.0Temperature Range: 37- 60°C(1)

Working Concentration: 0.05 - 1 mg/ml

Stabilizer: Ca2+; Activity decreases if Ca2+ is removed from the solution(2)

Activity: Active in the presence of 1% SDS, 4 M Urea(3), 1% Triton X-100(3), 5% Tween-20, or 3 M Guanidine HCl.

Inactivated by PMSF and diisopropyl phosphoro-fluoridate (DPF). Partially inactivated by EGTA. Not inactivated by EDTA, thiol reagents, or trypsin/chymotrypsin inhibitors(1)

Inactivation: Inactivate by heating at 95°C for 10 minutes, PMSF or DPF.

Purity:Tested for contaminating exonucleases, endonucle-ases, and ribonucleases.

Storage buffer:20 mM Tris-HCl (pH 7.4), 1.0 mM CaCl2, and 50% glycerol.

Unit definition:One unit is the amount of enzyme that liberates folin positive amino acids and peptides corresponding to 1 µmol tyrosine in 1 minute at 37°C using hemoglo-bin as a substrate.

Activity: ≥ 600 units/ml

Specific activity: ≥ 30 units/mg

Protein concentration: 20 mg/ml

Recommended reaction buffer: 50 mM Tris-HCl (pH 7.4) and 1.0 mM CaCl2 and 50% glycerol.

Note: Proteinase K has two binding sites for Ca2+ which act as a stabilizing factor of the enzyme. When Ca2+ is removed from the enzyme approxi-mately 80% of the catalytic activity is lost because of the structural change(2). However, residual Proteinase K activity is usually sufficient to degrade proteins commonly present during DNA/RNA isolation.


References:1. Ebeling, W., Hennrich, N., Klockow, M., Metz, H.,

Orth, H. D. and Lang, H. (1974) Eur. J. Biochem. 47, 91-97.

2. Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor, New York.

3. Hilz, H., Wiegers, U. and Adamietz, P. (1975) Eur. J. Biochem. 56, 103-108.

4. Roelcke, D. and Ulenbruck, G. (1969) Z. Med. Mikrobiot. und Immunol. 155, 156-170.

5. Brdiczka, D. and Krebs, W. (1973) Biochem. Biophys. Acta 297, 203-212.

6. Williamson, J., Greene, J., Cherif, S. and Milner-White, E. J. (1977) Biochem. J. 167, 731-737.

Proteinase K Solution

Molecular Biology Products & Kits | Proteinases

PurificationPlasmid Purification

Midi to Giga Preps

Spin Columns

Endotoxin-Free Preps

Medium Throughput

High Throughput

Yeast

BAC

DNA Cleanup

Gel Extraction

Genomic DNA Isolation

PCR Purification

RNA Purification

Sequencing Dye Cleanup

Histidine-Tagged Protein

Purification Kits & Resins

GST-Tagged Protein Purification


Purification | Table of Contents

Plasmid Purification – Midi to Giga PrepsPrepEase® Midi Plasmid Kit 140PrepEase Maxi Plasmid Kit 140PrepEase Giga Plasmid Kit 140

Plasmid Purification – Spin ColumnsPrepEase Quick MiniSpin Plasmid Kit 141PrepEase MiniSpin Plasmid Kit 141

Plasmid Purification – Endotoxin-Free PrepsPrepEase Endotoxin-Free Maxi Plasmid Kit 142

Plasmid Purification – Medium ThroughputPrepEase Plasmid 8-well Strip Kit 143PrepEase 8-well Strip Starter Kit for Vacuum 143PrepEase Vacuum Manifold 143

Plasmid Purification – High ThroughputPrepEase Plasmid 96-well Plate Kit 144PrepEase Plasmid 96-well Plate Core Kit 144

Plasmid Purification – YeastPrepEase Yeast Plasmid Isolation Kit 145

Plasmid Purification – BACPrepEase BAC Purification Kit 145

DNA CleanupPrepEase DNA Cleanup Kits 146

Gel ExtractionPrepEase Gel Extraction Kits 146

Genomic DNA IsolationPrepEase Genomic DNA Isolation Kits 147

PCR PurificationPrepEase PCR Purification 96-Well Plate Kits

(Ultrafiltration) 148ExoSAP-IT® PCR Product Cleanup 148HT ExoSAP-IT High-Throughput PCR Product

Cleanup 149

RNA PurificationPrepEase RNA Spin Kits 150PrepEase mRNA MiniSpin Kit 150PrepEase RNA/Protein Spin Kit 151

Sequencing Dye CleanupPrepEase Sequencing Dye Cleanup Kits 152

Histidine-Tagged Protein Purification KitsPrepEase Histidine-Tagged Mini Kit – High

Specificity 153PrepEase Histidine-Tagged Midi Kit – High

Specificity 153PrepEase Histidine-Tagged Maxi Kit – High

Specificity 153PrepEase Histidine-Tagged Mini Kit – High

Yield 154PrepEase Histidine-Tagged Midi Kit – High

Yield 154PrepEase Histidine-Tagged Maxi Kit – High

Yield 154

Histidine-Tagged Protein Purification ResinsPrepEase Histidine-Tagged High Specificity

Purification Resin 155PrepEase Histidine-Tagged High Yield

Purification Resin 155

GST-Tagged Protein PurificationPrepEase Protein Purification Glutathione

Agarose 4B 155


Purification | Plasmid Purification – Midi to Giga Preps

PrepEase Midi Plasmid KitBinding capacity up to 100 µgCulture volume up to 30 ml78706 100 preps

PrepEase Maxi Plasmid KitBinding capacity up to 500 µgCulture volume up to 150 ml78711 50 preps

PrepEase Giga Plasmid KitBinding capacity up to 10 mgCulture volume up to 2,000 ml78720 5 preps

Designed to obtain low to high copy plasmid DNA from bacterial cultures.

�� Based on familiar anion-exchange columns �� For routine purification using a well-characterized method �� For use with plasmids from 3 - 10 kb�� Purify from 100 µg up to 10 mg depending on the column size�� Greater than 90% recovery of ultrapure plasmid

PrepEase Plasmid Purification Kits employ a modi-fied alkaline–SDS lysis procedure with patented anion-exchange columns to isolate high purity plasmid DNA from bacterial cell lysates. During the cell lysis step, both chromosomal and plasmid DNA are denatured. Potassium acetate is added to form a neutralized precipitate containing chromo-somal DNA and other cellular components. Plasmid DNA remains in the solution, reverts to its native supercoiled structure, and is then loaded onto an equilibrated PrepEase Column. The plasmid DNA becomes bound to the anion-exchange resin and is then eluted from the column with washing steps. Eluted DNA is precipitated and easily dissolved in TE Buffer or nuclease-free water. The purified plasmids are suitable for use in the most demanding molecu-lar biology applications, including transfection, in vitro transcription, automated or manual sequencing, cloning, hybridization, and PCR.

PrepEase Columns are stable over a wide pH range, from pH 2.5 - 8.5, and can remain in contact with buffers for up to three hours without any change in chromatographic properties. After three hours, nucleic acids will begin to elute at increasingly lower salt concentrations. Normally, the resin remains functional in buffers containing up to 2 M salt, in denaturing agents such as formamide, urea, or in common detergents such as Triton® X-100 and Igepal™ (NP-40 substitute). The protocols are suit-able for purifying most plasmids ranging from 3 to 10 kb. Two protocols are presented, one for purifying high-copy number plasmids (> 20 copies/cell) and the other for purifying low-copy number plasmids (< 20 copies/cell).

Properties of PrepEase ResinPrepEase Resin is a silica-based anion-exchange resin, used for routine separation of different types of nucleic acids. Patented PrepEase Resin forms the basis for the entire line of USB nucleic acid purifica-tion products. The resin consists of hydrophilic, macro-porous silica beads coupled to a methyl-ethylamine functional group. The functional group provides a high overall charge density that permits the negatively charged plasmid DNA to bind with high specificity to the resin.

The beads are uniform in diameter due to a specialized manufacturing process that is rigorously controlled and monitored, and contain particularly large pores. The special properties of PrepEase Resin allow for optimum flow rates through the column and more efficient binding of nucleic acids to the matrix.

Kit components:Each kit includes PrepEase gravity-flow columns and all the necessary reagents for ultrapure plasmid purification.

Includes specialized PrepEase Folded Filters to remove cellular debris from lysates.

– Gentle treatment – No shearing of DNA – Fast procedure

Shipping and storage:Shipped ambient. Store at room temperature.

Related products

PrepEase Endotoxin-free Maxi Plasmid Kit78730 10 preps

PrepEase MiniSpin Plasmid Kit78737 250 preps

PrepEase Plasmid 8-well Strip Kit78745 12 strips

PrepEase Plasmid 96-well Plate Kit78751 4 x 96-well plates


PrepEase Plasmid Gravity-Flow Column Kits


78742 250 preps

Very rapid purification

�� Rapid processing of 1 - 3 ml of culture �� For purifying vectors up to 15 kb �� Yields up to 15 μg �� Ideal for direct sequencing �� 50 μl elution volume �� Up to 95% recovery

The PrepEase Quick MiniSpin Plasmid Kit enables rapid, simple purification of plasmid DNA from small volumes of bacterial culture. The kits employs an alkaline lysis procedure and spin columns containing special, patented, anion-exchange membrane filters, which save time by eliminating extra wash and drying steps.

The PrepEase Quick MiniSpin Plasmid Kit protocol is designed for preparation of plasmids ranging up to 15 kb in size, and for high-copy plasmids in particular. Simple modifications enable use of a wide

variety of bacterial strains and growth media, and optimization of elution procedures. For preparation of low-copy plasmids, try the USB PrepEase MiniSpin Plasmid Kit.

Kit components:Includes PrepEase Spin Columns, collection tubes, and all the necessary reagents for rapid plasmid purification.


PrepEase Quick MiniSpin Plasmid Kit

Purification | Plasmid Purification – Spin Columns

78737 250 preps

High yield purification

�� Process up to 10 ml of culture �� For purifying vectors up to 15 kb �� Yields up to 40 μg �� Ideal for multiple analyses �� 50 μl elution volume �� Up to 95% recovery

The PrepEase MiniSpin Plasmid Kit enables rapid, simple purification of plasmid DNA from small volumes of bacterial culture. The kit employs an al-kaline lysis procedure and patented, anion-exchange membrane filter spin-columns.

The PrepEase MiniSpin Plasmid Kit is designed for preparation of plasmids ranging up to 15 kb in size. Simple modifications of the protocol enable use of a wide variety of bacterial strains and growth media, elimination of problematic nucleases, optimization of wash and elution procedures, and purification of low-copy plasmids. For even faster preparation of high-copy plasmids, try the USB PrepEase Quick MiniSpin Plasmid Kit.

Kit components:Includes PrepEase Spin Columns, collection tubes, and all the necessary reagents for rapid plasmid purification.


PrepEase MiniSpin Plasmid Kit

PrepEase Standard MiniSpin Column.

PrepEase Quick MiniSpin Column.

Fig. 1. The PrepEase Quick MiniSpin Kit gives high-quality results for auto-mated sequencing. Typical result from plasmid DNA purified using the PrepEase Quick MiniSpin Plasmid Kit, sequenced and analyzed on an ABI PRISM® 3700. Please note the absence of “no calls (N)” far behind the reading length of 700 bp.


Binding capacity up to 500 µgCulture volume up to 150 ml78730 10 preps

Designed to obtain endotoxin-free plasmids for transfections

�� Based on familiar anion-exchange columns�� Yields plasmid with less than 0.05 EU/µg DNA�� Purify from 500 µg up to 10 mg depending on the column size�� Greater than 90% recovery of ultrapure plasmid

Endotoxins (lipopolysaccharide, LPS) are a major component of gram-negative bacterial cell walls and are extremely potent stimulators of the mammalian immune system. LPS is a common contaminant of plasmid DNA preparations grown in E. coli. The negative charges associated with lipid A and the inner core of LPS cause these molecules to behave like DNA on anion-exchange chromatography resins. Therefore, standard column chromatography procedures typically yield slightly contaminated plasmid DNA.

The PrepEase Endotoxin-Free (EF) Maxi Plasmid Kit utilizes a modified alkaline/SDS lysis procedure to prepare the bacterial cell pellet for plasmid purifica-tion. Both chromosomal and plasmid DNA are dena-tured under alkaline conditions. Potassium acetate is then added to the denatured lysate to form a precipitant of chromosomal DNA and other cellular compounds. The potassium acetate buffer also neu-tralizes the lysate. Plasmid DNA remains in solution and reverts to its native supercoiled structure. The plasmid DNA is bound to an equilibrated PrepEase-EF Column and then eluted after efficient washing of the column. Endotoxins are removed by using a specialized solution, N3-EF Buffer. After precipitation of the eluted DNA, it can easily be dissolved in TE-EF Buffer or EF-Water for further use.

Properties of PrepEase ResinPrepEase-EF Columns contain anion exchange silica resin packed between two inert filter elements. The columns may be used over a wide pH range, from pH 2.5 - 8.5, and can remain in contact with buffers for up to three hours without any change in chromatographic properties. After three hours, nucleic acids will begin to elute at increasingly lower salt concentrations. Normally, the resin remains functional in buffers containing up to 2 M salt. In addition, the resin remains intact in the presence of denaturing agents such as formamide, urea, or common detergents such as Triton X-100 and Igepal (NP-40 substitute).

Kit components:Each kit includes PrepEase Endotoxin-Free Gravity-Flow Columns and all the necessary reagents for ultrapure plasmid purification.

Includes specialized PrepEase Folded Filters/Bottle Top Filters to remove cellular debris from lysates.

– Gentle treatment – No shearing of vectors or constructs– Fast procedure


Related products

Agarose - Separation ≥ 500 bpGenetic Performance Certified™75817 100 gm 250 gm

Agarose - LE 32802 100 gm 250 gm

Ampicillin, Sodium Salt 11259 5 gm 25 gm

Chloramphenicol23660 25 gm

Kanamycin Sulfate 17924 5 gm 25 gm

LB Agar, Ready-Made Powder 75851 250 gm

1 kg

LB Broth, Ready-Made Powder 75852 250 gm

1 kg

Lysozyme 18645 5 gm 10 gm

25 gm

RapidRun™ Agarose Buffer, 20X Solution 77523 1 L

5 L

Streptomycin Sulfate 21865 10 gm

100 gm


5 L

TBE Buffer, 5X Solution 75891 1 L

5 L

TBE Buffer, 10X Ready-Mixed Powder70454 6 x 200 ml

TE Buffer, 1X Solution75893 10 x 1 ml

100 ml 500 ml

Tetracycline Hydrochloride 22105 25 gm


100 ml

PrepEase Endotoxin-free Maxi Plasmid Kit

Purification | Plasmid Purification – Endotoxin-free Preps

Fig. 1. Typical results with PrepEase plasmid purification products.


Binding capacity up to 20 µgCulture volume up to 5 ml 78745 12 strips

Designed for the isolation of high copy plasmid DNA using 8 well strips

�� Based on familiar silica-membrane technology�� Time-saving purification of multiple samples using 8-well strips�� For use with plasmids up to 15 kb�� Yields up to 20 μg highly pure plasmid DNA from 5 ml of culture: 5 - 15 μg from 1.5 ml of culture�� Process samples by vacuum or robotics

The PrepEase Plasmid 8-well Strip Kit enables rapid purification of plasmid DNA from small volumes of bacterial culture. The kit employs an alkaline lysis procedure, followed by either vacuum filtration, or optionally by robotics, with easy-to-use filter and plasmid binding strips. The plasmid binding strips contain patented anion-exchange membrane filters, allowing highly specific purification of plasmid DNA.

Plasmids ranging in size up to 15 kb may be purified with yields up to 15 μg from 1.5 ml of bacterial culture. The kit is suited for high copy number plasmid purification. Low copy plasmids may be puri-fied by increasing the culture volumes to 5.0 ml. Simple modifications of the protocol enable use of a wide variety of bacterial strains and growth media, elimination of problematic nucleases, and optimization of washes to increase yield.

Use of the PrepEase Plasmid 8-well Strip Kit for the first time requires additional accessories, available separately. For standard vacuum processing, use the PrepEase Vacuum Manifold (PN 78780) and the PrepEase 8-well Strip Starter Kit for Vacuum (PN 78784).

Kit components:Kit includes PrepEase 8-well Binding Strips and all the necessary reagents for high purity plasmid purification.

Includes specialized PrepEase Filter Strips to clear bacterial lysates–replaces time-consuming centrifugation.


Related products

PrepEase 8-well Strip Starter Kit forVacuumIncludes 2 x 8-well strip holders, dummy strips to close unused rows, and accessories for processing manually or on robotic platform.78784 1 pack

PrepEase Vacuum ManifoldFor use with PrepEase 8-well strips and 96-well plates. Consists of manifold base and lid, spacer set, and a waste tray.78780 1 ea

PrepEase Plasmid 8-well Strip Kit

Purification | Plasmid Purification – Medium Throughput

PrepEase 8-Well Strip Procedure


Binding capacity up to 20 µgCulture volume up to 5 ml 78751 4 x 96-well plates

Designed for the isolation of high copy plasmid DNA using 96 well plates

�� Based on familiar silica-membrane technology �� Time-saving purification of multiple samples using 96-well plates �� For use with plasmids up to 15 kb �� Yields up to 20 µg highly pure plasmid DNA from 5 ml of culture; 5 - 15 µg from 1.5 ml of culture

The PrepEase Plasmid 96-well Plate Kit enables rapid, simple purification of plasmid DNA from small volumes of bacterial cultures. The kit uses an alkaline lysis procedure, followed by vacuum filtration (and optionally, centrifugation) with filter and plasmid binding plates. The plasmid binding plates contain patented anion-exchange membrane filters, allowing highly specific purification of plasmid DNA. The PrepEase Plasmid 96-well Plate Kit requires a vacuum manifold, such as the PrepEase Vacuum Manifold (PN 78780) or the QiaVac™ 96.

Plasmids ranging up to 15 kb in size may be purified with yields of up to 15 µg from 1.5 ml of bacte-rial culture. The kit is suited for high copy number plasmid purification. Low copy plasmids may be purified by increasing the culture volumes to 5.0 ml. Simple modifications of the protocol enable use of a wide variety of bacterial strains and growth media, elimination of problematic nucleases and optimiza-tion of washes to increase yield.

Kit components:Includes square-well blocks and cover sheets for bacterial cultivation, filter plates, binding plates, wash plates and buffers


Related products

PrepEase Plasmid 96-well Plate Core KitIncludes filter plates, binding plates, wash plates and buffers. Does not contain square-well blocks for bacterial cultivation.78753 24 x 96-well plates

Related products – Ultrapure reagents

Agarose - Separation ≥ 500 bpGenetic Performance Certified75817 100 gm 250 gm

Agarose - LE 32802 100 gm 250 gm

LB Agar, Ready-Made Powder 75851 250 gm 1 kg

LB Broth, Ready-Made Powder 75852 250 gm

1 kg

Lysozyme 18645 5 gm 10 gm

25 gm

RapidRun Agarose Buffer, 20X Solution 77523 1 L

5 L


5 L

TBE Buffer, 5X Solution 75891 1 L

5 L


TE Buffer, 1X Solution75893 10 x 1 ml

100 ml 500 ml

Water, Nuclease-Free 71786 10 x 1 ml 100 ml

Related products – Ultrapure antibiotics

Ampicillin, Sodium Salt 11259 5 gm 25 gm

Chloramphenicol23660 25 gm

Kanamycin Sulfate 17924 5 gm 25 gm

Streptomycin Sulfate 21865 10 gm

Tetracycline Hydrochloride 22105 25 gm

PrepEase Plasmid 96-well Plate Kit

Purification | Plasmid Purification – High Throughput

PrepEase Plasmid Filter Plate

PrepEase Plasmid Binding Plate vacuum

vacuum

highly pure plasmid DNA

screening

restrictiondigests

sequencing

cloning


79220 50 preps

The PrepEase Yeast Plasmid Isolation Kit is designed for the isolation of 2 µ plasmids from either yeast patches on plates or from yeast grown in small liquid culture. The isolation protocol eliminates the need for phenol/chloro form extraction, glass bead disruption and column elution.

The isolated plasmid DNA can be used for PCR amplification, bacterial transformation or for direct sequencing. The ability to sequence directly from the isolated plasmid without the need for bacterial transfor mation or a pre-amplification step and restriction enzyme digestion makes it an efficient method for screening yeast transformants. The kit may also be used to obtain spheroplasts for other analysis.

Direct purificationMinimize the time required to analyze yeast plas-mids, e.g. yeast two-hybrid experiments. The need to shuttle plasmids from yeast to E. coli is eliminated.

Simple protocolAllows the isolation of plasmid from yeast cultures almost as rapidly and readily as from E. coli.

SafeNo organic extraction or mechanical disruption of yeast samples is involved.

Quality resultsYields plasmid DNA suitable for a variety of applica-tions, including direct sequencing, PCR amplifica-tion, and transformation of chemically competent bacterial cells.

Kit components:Enzyme Solution Spheroplast SolutionLysis SolutionPrecipitation Solution


PrepEase Yeast Plasmid Isolation Kit

Binding capacity up to 100 µgCulture volume up to 150 - 500 ml 78722 10 preps

Designed for purification of BACs, PACs, P1s, and other large constructs

�� Based on familiar anion-exchange chromatography�� Purify constructs up to 300 kb�� Yield is typically 10 - 100 µg of BAC vector

The PrepEase BAC Purification Kit employs a modi-fied alkaline–SDS lysis procedure with patented anion-exchange columns to isolate high purity plasmid DNA from bacterial cell lysates. During the cell lysis step, both chromosomal and plasmid DNA are denatured. Potassium acetate is added to form a neutralized precipitate containing chromo-somal DNA and other cellular components. Plasmid DNA remains in the solution, reverts to its native supercoiled structure, and is then loaded onto an equilibrated PrepEase Column. The plasmid DNA becomes bound to the anion-exchange resin and is then eluted from the column with washing steps. Eluted DNA is precipitated and easily dissolved in TE Buffer or nuclease-free water. The purified plasmids are suitable for use in the most demanding molecu-lar biology applications, including transfection, in vitro transcription, automated or manual sequencing, cloning, hybridization, and PCR.

PrepEase Columns contain anion exchange silica resin. The columns are stable over a wide pH range, from pH 2.5 - 8.5, and can remain in contact with buffers for up to three hours without any change in chromatographic properties. After three hours, nucleic acids will begin to elute at increasingly lower salt concentrations. Normally, the resin remains functional in buffers containing up to 2 M salt, in denaturing agents such as formamide, urea, or in common detergents such as Triton X-100 and Igepal (NP-40 substitute).

The BAC purification (low-copy) protocol is suitable for the isolation of low copy plasmids and contains enough buffers and RNase A to perform 10 preps. Typical yields are 10 - 100 µg from 500 ml of fermentation broth depending on copy number and size of constructs.

Kit components:Kit includes PrepEase Gravity-Flow Columns and all the necessary reagents for ultrapure large construct purification.

Specialized PrepEase Folded Filters are used to remove cellular debris from lysates (Fig. 1). – Gentle treatment – No shearing of vectors or constructs – Fast procedure


PrepEase BAC Purification Kit

Purification | Plasmid Purification – Yeast/BAC

Fig. 2. Typical result using the PrepEase BAC Purification Kit.

Fig. 1. PrepEase Folded Filters are used to remove SDS and cellular debris from bacterial lysates and eliminate a cen-trifugation step.


Purification | DNA Cleanup/Gel Extraction

Binding capacity up to 15 µg78756 50 preps78757 250 preps

Designed for purifying DNA fragments from agarose gels, and also suited for PCR cleanup

�� Spin columns based on familiar silica-membrane technology�� High recovery rates of up to 90% with low elution volume (15 µl)�� Purify fragments from 50 bp - 10 kb�� Ideal for purifying restriction-digested DNA in gels or cleanup of reaction products

The PrepEase Gel Extraction Kit may be used to purify DNA from agarose gels or to cleanup reaction products. The kit may be used for desalination, re-moval of enzymes, nucleotides, or labeling reagents from sample DNA.

With the PrepEase Cleanup method, DNA binds in the presence of chaotropic salts to the silica mem-brane within the spin column. Contaminants such as salts and soluble macromolecular components are removed with a simple washing step. Pure DNA is eluted under low ionic strength conditions with slightly alkaline buffer.

Typical recovery rates range from 70 - 95% for DNA fragments between 50 - 10,000 bp. The purified DNA may be used directly in applications such as DNA sequencing, PCR, cloning, or probe labeling.

Kit components:Includes PrepEase Spin Columns and all the neces-sary reagents for rapid DNA purification.


Related products – Ultrapure reagents

Agarose - LE 32802 100 gm

250 gm

Agarose - Low Melt, Separation ≥ 1000 bpGenetic Performance Certified 32830 25 gm

100 gm

Agarose - Low Melt, Separation ≤ 1000 bpGenetic Performance Certified32829 25 gm

100 gm

Ethidium Bromide Drops75816 5 ml


5 L

Related products – Markers and ladders


DNA Ladder, 100 bp76712 500 µl

PCR Markers, 50 – 2000 bp76710 250 µl

PrepEase Gel Extraction Kits

washing

elution

binding

solubilized gel slice

+ NT Buffer

agarose gel

highly pure DNA fragments

ligation

sequencing

labeling

restrictiondigests


Designed for the purification of DNA from reactions or for buffer exchange

�� Spin columns based on familiar silica-membrane technology�� High recovery rates up to 90% with low elution volume (15 μl)�� Ideal for cleanup of reaction products�� Binding capacity up to 15 μg�� For DNA fragments 50 – 10,000 bp

The PrepEase DNA Cleanup Kit is designed to purify DNA fragments from enzymatic reactions or for buffer exchange. Thus, this kit can be used to remove

contaminants such as salts, enzymes, nucleotides, labeling reagents, etc. from a DNA sample. This kit can also be used for purification of DNA from Chromatin Immunoprecipitation (ChIP) assays.

The kit provides a simple and convenient way of purifying DNA by selective binding to a silica membrane within the spin column in the presence of a chaotropic salt. The membrane is then washed to remove contaminants such as salts and soluble macromolecules. Pure DNA is then eluted from the membrane with buffer.

Each kit includes PrepEase Cleanup Columns, collection tubes and the necessary reagents for the purification of DNA fragments.

Storage:Ships ambient. Store at room temperature.

PrepEase DNA Cleanup Kits

binding

washing

elution


Purification | Genomic DNA Isolation

78850 50 preps78855 250 preps

Designed for the rapid isolation of genomic DNA

�� Does not use phenol or columns�� Processes large number of samples in a short time�� Optimized for maximal extraction of high molecular weight DNA�� Yields up to 90 μg per prep depending on sample type

The PrepEase Genomic DNA Isolation Kit provides a simple and efficient procedure for the rapid isolation of genomic DNA from various samples, including mouse tails, animal tissues, cultured cells, bacteria, yeast, whole blood, and plant leaves. The procedure does not use phenol or columns and can process a large number of samples in a short time.

This kit has been optimized for maximal extraction of pure high molecular weight genomic DNA. Such genomic DNA is free of protein and ideal for down-stream analyses, including restriction endonuclease digestion, PCR, library construction, Southern blot-ting, cloning, and other applications. The specially formulated Homogenization Buffer not only works well with tissues containing high amounts of nucle-ases, but also with tough fibrous tissues.

Each kit includes all the necessary reagents for the isolation of genomic DNA from a variety of samples. Proteinase K is included for treatment of skin, mouse tails, and other hard tissues.

Storage:Ships ambient. Store individual components as specified.

PrepEase Genomic DNA Isolation Kits

Fig. 1. PCR analysis of transgenic mice. Genomic DNA isolated from mouse tails was analyzed by multiplex PCR to detect three transgenes. PCR products were resolved by electrophoresis in a 1% agarose gel. The unique product for each transgene is shown at the left side of the panel. The mouse genotype is indicated at the bottom of the figure. (Image courtesy of Jianqi Yang, Case Western Reserve University.)

Fig. 3. Genomic DNA isolated from various samples. Genomic DNA was resolved by electrophoresis in a 0.7% agarose gel. Samples are from mouse unless otherwise specified. (Image courtesy of Jianqi Yang, Case Western Reserve University.)

Fig. 2. Southern blotting of C/EBPα alleles in several tissue types. Genomic DNA isolated from various mouse tissues was ana-lyzed using Southern blotting. The unique band for each C/EBPα allele is shown at the left side of the panel; the name of tissue is indicated at the right side. Each lane represents DNA isolated from a single ani-mal. (Image courtesy of Jianqi Yang, Case Western Reserve University.)


78250 20 reactions78200 100 reactions78201 500 reactions78202 2,000 reactions78205 5,000 reactionsCustom formulations by request

�� Conserves PCR samples – 100% recovery of both short and long PCR products �� One tube/one-step PCR cleanup – Add ExoSAP-IT reagent directly to PCR product.�� Eliminates spin columns – Decreases time and expense while increasing yield�� Removes contaminating primers and dNTPs – No interference in downstream applications�� Scalable – Economical for high-throughput purification�� Simple processing – Robotic-friendly; Replaces beads, filtrations, and plates�� Generates less waste than columns

ExoSAP-IT reagent is designed for simple, quick PCR cleanup for downstream applications, such as DNA sequencing or Single Nucleotide Polymor-phism (SNP) analysis. When PCR amplification is complete, any unused dNTPs and primers remain-ing in the PCR product mixture will interfere with these methods. ExoSAP-IT PCR Cleanup removes these contaminants.

ExoSAP-IT reagent is added directly to the PCR product and incubated at 37°C for 15 minutes. After PCR treatment, it is inactivated simply by heating to 80°C for 15 minutes (Fig. 1).

ExoSAP-IT single-step PCR cleanup utilizes two hydrolytic enzymes, Exonuclease I and Shrimp Alkaline Phosphatase (SAP), together in a specially formulated buffer, to remove unwanted dNTPs and primers from PCR products. Exonuclease I removes residual single-stranded primers and any extraneous single-stranded DNA produced in the PCR. SAP re-moves the remaining dNTPs from the PCR mixture.

Rapid PCR product cleanup protocolThe ExoSAP-IT method requires only one pipet-ting step and two incubations. Just add ExoSAP-IT reagent to the PCR product and within 30 minutes sequencing or SNP analysis can be performed.

Simple: single-stepThe method is designed to require a minimum of “hands-on” time. Enzymatic removal of excess primers and unincorporated nucleotides occurs in one easy step by using ExoSAP-IT reagent in a single tube or microtiter well. Only simple pipette transfers are required, therefore, many samples can be pro-cessed at once, either manually or with robotics.

No sample lossUse of ExoSAP-IT reagent eliminates all gel or column purifications, sedimentations, filtrations, beads, and/or magnetic separations(1). There is 100% recovery of both short and long PCR products with ExoSAP-IT reagent.

Achieve high quality data from PCR productsExoSAP-IT reagent may be used as an effective cleanup method prior to fluorescent or radioac-tive DNA sequencing, SNP analysis or any other application requiring a PCR product free of excess nucleotides and primers.








Related product


ExoSAP-IT PCR Product Cleanup

Purification | PCR Purification

78761 10 x 96-well plates78762 50 x 96-well plates

Designed for high throughput purification of PCR products using 96-well plates

�� Ultrafiltration technology �� For PCR products ≥150 bp �� Removes contaminating enzymes, salts, primers, and dNTPs �� Process 20 - 300 μl PCR volume per well �� Recover up to 95% depending on length of PCR product in as little as 25 μl

May be performed manually by vacuum, by centrifu-gation, or by using an automated workstation.

Kit components:Includes 96-well plates and buffer for PCR product purification.


PrepEase PCR Purification 96-Well Plate Kits (Ultrafiltration)

PCR Mixture Post-Amplification

PCR Product

+

Nucleosides

+

Inorganic Phosphate (Pi)

Add ExoSAP-IT®

Excess Primers

Excess dNTPs

37°C, 15 min for treatment80°C, 15 min to inactivate

Fig. 1. Summary of ExoSAP-IT PCR product treatment.


Purification | PCR Purification

78395 480 rctns x 8-tube strip5,760 rctns x 1 plate

(12 x 8-tube strips)23,040 rctns - 4 plates

1,000 rctns (2 ml) 5,000 rctns (10 ml)

�� One-tube PCR cleanup – Add HT ExoSAP-IT reagent directly to PCR product.�� Exceptional accuracy – Achieve high quality data even with long read lengths (Fig. 1)�� 100% sample recovery – No loss of PCR products regardless of the fragment size (Fig. 2)�� Simple, single-step – Replace multiple steps and wasted time with bead or column use�� High-throughput processing – Even faster time to results with a low viscosity formulation, allowing robotic pipetting�� Removes excess primers and dNTPs – Does not interfere with downstream applications • sequencing • SNP analysis • single base extension • fragment analysis • in vitro transcription�� Scalable – Treat reaction volumes from 5 µl to 5 L�� Convenient packaging – Available in 8-tube strips and 12 x 8-tube strips in a 96-well plate �� Stable at +25°C for 8 hours – Retains full functional activity and at 4°C is stable for one week (Fig. 3)

HT ExoSAP-IT High-Throughput PCR Product Cleanup is an alternative formulation of ExoSAP-IT reagent specifically designed for the unique requirements of high-throughput, automated platform and multi-channel pipettes. HT ExoSAP-IT reagent has both a longer lifetime at higher tempera-tures and a decreased viscosity for robotic pipetting. Like ExoSAP-IT PCR Product Cleanup (PN 78200), HT ExoSAP-IT reagent is a mixture of Exonuclease I and Shrimp Alkaline Phosphatase (SAP) which removes excess primers and dNTPs following a PCR reaction

in a single incubation. HT ExoSAP-IT reagent possesses greater thermal stability and lower viscosity which provides greater ease-of-use in automated 96- and 384-well formats.

HT ExoSAP-IT reagent is added directly to the PCR product and incubated at 37°C for 15 minutes. After PCR treatment, it is inactivated simply by heating to 80°C for 15 minutes. Our high-throughput formula-tion is designed for robotic pipetting and multi-channel pipettes. HT ExoSAP-IT reagent has a lower viscosity than the standard mix and is available in an 8-tube strip format for ease of use.

HT ExoSAP-IT reagent utilizes two hydrolytic enzymes, Exonuclease I and SAP, together in a specially formulated buffer, to remove unwanted dNTPs and primers from PCR products. Exonuclease I removes residual single-stranded primers and any extraneous single-stranded DNA produced in the PCR. SAP removes the remaining dNTPs from the PCR mixture.

Simple: Single-stepHT ExoSAP-IT reagent requires only one pipetting step and two incubations. Just add HT ExoSAP-IT reagent to the PCR product and within 30 minutes sequencing, SNP analysis, single base extension, or fragment analysis can be performed.

Enzymatic removal of excess primers and unincorpo-rated nucleotides occurs in one easy step by using HT ExoSAP-IT reagent in a single-tube, 8-tube strip, or 12 strips in a 96-well low skirted PCR plate. Our new formatting makes this an ideal product for use with robotics.

Exceptional accuracy with HT ExoSAP-IT Achieve high data quality and sequencing accuracy with HT ExoSAP-IT reagent, even with long read lengths. Sequence reads of PCR products treated with HT ExoSAP-IT reagent were on average 50+ bases longer than samples treated with competitor products. Using human genomic DNA, a 1,007 bp fragment was sequenced with no miscalls (Fig. 1). The size limitations associated with alternative PCR cleanup methods are not a factor with HT ExoSAP-IT reagent. Phred 20 values over the entire length of a 970 bp sequence was 822 ± 9 with HT ExoSAP-IT-treated samples and only 776 ± 83 with samples treated with Agencourt® AMPure® XP (see Table 1).

Phred score values of samples processed with HT ExoSAP-IT and AMPure XP BeadsTable 1. HT ExoSAP-IT reagent gives better sequencing results.

HT ExoSAP-IT

AMPure XP beads

Number bases read (Average of 16 samples) 901 850

Total miscalls 0 25Pass rate (%)* 100% 99.7%Phred 20** 822 ± 9 776 ± 83

* Pass rate – Average Phred value greater than 20 bases between 100 bp and 300 bp.

** Phred 20 values over the entire length of sequence (no trim-ming ~970 bp)

No sample lossUse of HT ExoSAP-IT eliminates all gel or column purifications, sedimentations, filtrations, beads, and/or magnetic separations(1). There is 100% recovery of both short and long PCR products with HT ExoSAP-IT High-Throughput PCR Product Cleanup (Fig. 2).

HT ExoSAP-IT-treated PCR product is stableHT ExoSAP-IT reagent is rigorously tested and subject to strict quality control. Fig. 3 demonstrates that no product degradation occurred after storage of HT ExoSAP-IT-treated PCR product for one week at 25°C.


References:See references on previous page (PN 78200).

HT ExoSAP-IT High-Throughput PCR Product Cleanup

HT ExoSAP-IT

AMPure XP

3 miscalls

Fig. 1. Sequencing of a 1,007 bp treated PCR product. A 1,007 bp fragment was amplified and treated with HT ExoSAP-IT reagent (above) or Agencourt AMPure XP beads (below) and sequenced. Pherograms revealed no miscalls with HT ExoSAP-IT but three miscalls with Agencourt AMPure XP beads at position 203, 204, and 220.

Fig. 2. Greater recovery of PCR product with HT ExoSAP-IT reagent. Equivalent volumes of PCR product were visualized on an ethid-ium bromide agarose gel and the band volume determined using image analysis. HT-ExoSAP-IT reagent allowed recovery of 100% of the 86 and 103 bp PCR products but AMPure XP beads allowed only 8 and 14% recovery, respectively. Un – Untreated, HT – HT-ExoSAP-IT treated, XP – AMPure XP purified, 100 bp – Affymetrix 100 bp ladder, bands from 100 to 1,000 bp by 100 bp intervals (PN 76712).

Fig. 3. High stability of HT ExoSAP-IT-treated products. A 1,007 bp DNA fragment amplified from genomic DNA with HotStart FideliTaq Master Mix (PN 71156) was treated with HT ExoSAP-IT reagent and stored for seven days at: -20ºC (lane 1), 4ºC (lane 2), or 25ºC (lane 3). Samples were visualized on 1.5% agarose/TAE/ethid-ium bromide gel. Lane MW: DNA Ladder, 1 kb Plus (PN 76714).

1 kb -20°C 4°C 25°C

86 bp 103 bp100 bp Un HT XP Un HT XP

545 bp 1,007 bp100 bp Un HT XP Un HT XP


Purification | RNA Purification

78878 12 preps

Designed to purify high quality poly(A) RNA from total RNA samples using PrepEase Oligo(dT) Latex Beads.

�� Yields up to 10 µg poly(A) RNA from up to 250 µg total RNA�� Exceptional yield – recover >90% of mRNA from starting total RNA sample�� Fast protocol - process 6 samples in less than 30 minutes�� Excellent purity - purify mRNA from previously isolated total RNA

The PrepEase mRNA MiniSpin Kit produces purified mRNA that is immediately ready for use in demand-ing downstream applications such as qRT-PCR and microarray analysis.

Kit components:Includes PrepEase Oligo(dT) Latex Beads, microfil-ters, and all the necessary reagents for isolation of poly(A) RNA from total RNA samples.

Shipping and storage: Ships ambient. Store PrepEase Oligo(dT) Latex Beads at 4°C. All other kit components may be stored at room temperature.

PrepEase mRNA MiniSpin Kit

Fig. 1. Poly(A) RNA yield vs. total RNA input

Fig. 2. Percent recovery of mRNA (GAPDH) and reduction of ribosomal RNA (rRNA) by PrepEase mRNA MiniSpin Kit from HeLa total RNA. Samples were analyzed by two-step RT-RCR (PN 75780 and 75762).


Designed for the isolation of total RNA from cells and tissue

�� Spin columns based on familiar silica membrane technology�� Less than 30 minutes hands-on time�� Binding capacity up to 100 µg�� Yields up to 70 µg of high purity DNA-free RNA�� DNase I is included for on-column digestion of contaminating genomic DNA�� Purified RNA may be used directly for RT-PCR, Northerns, microarrays, or cDNA synthesis

Purify total RNA from: Up to 5 x 106 cultured mammalian cells 0.5 - 30 mg mammalian tissue Up to 109 bacterial cells Up to 108 yeast cells Up to 100 µl biological fluids (e.g. serum) RNA cleanup from reaction mixtures

PrepEase RNA Spin Kits are ideal for the isolation of total RNA from mammalian cells, tissues and yeast. The purified RNA has an A260/280 ratio generally ex-ceeding 1.9 and is ready for use in applications such as RT-PCR, primer extension, microarrays, or RNase protection assays. Even biological samples, which are sometimes difficult to process, will yield high quality RNA with the PrepEase RNA Spin Kit. The standard protocol yields up to 70 µg of total RNA per PrepEase RNA Spin Column from up to 5 x 106 cultured cells or 30 mg of tissue.

Kit components:Includes PrepEase Filters to reduce lysate viscosity.

Includes PrepEase RNA Spin Columns, collection tubes, and all the necessary reagents for ultrapure RNA purification.


PrepEase RNA Spin Kits

cell lysis

homogenization by filtration

binding

DNase I incubationdirectly on thecolumnwashing

elution

Northern blotting primer extensionarray technology

RT-PCR RNase protection

PrepEase RNA and Plant RNA Spin Procedure


Purification | RNA Purification

78871 50 preps

Designed for high quality purification of total RNA and protein from the same sample to allow easy comparison of RNA and protein expression

�� High purity RNA – on column DNase digestion to remove contaminating DNA�� Total protein is prepared in loading buffer, ready for SDS-PAGE analysis�� Straightforward protocol allows efficient workflow

The PrepEase RNA/Protein Spin Kit provides purified total RNA that is immediately ready for sensitive downstream applications such as qRT-PCR and northern blotting. Total protein is eluted in loading buffer for SDS-PAGE analysis.

This kit yields up to 70 μg total RNA and 1.2 mg total protein from a wide array of starting materials, including mammalian tissues and cells, bacteria, yeast, and biological fluids.

Kit components:Includes PrepEase RNA & Protein Spin Columns, filter units, collecting tubes, and all the reagents necessary to isolate both total RNA and protein from the same starting sample.

Shipping and storage: Ships ambient. Store lyophilized rDNase and TCEP at 4°C upon receipt. All other kit components can be stored at room temperature (20-25°C).

PrepEase RNA/Protein Spin Kit

Fig. 1. Procedure overview for the purification of RNA and protein from one sample.

Proteinwash

Proteinprecipitation

RNA washes

RNA elution

rDNase digest

Homogenize sample

Lyse cells

Filter lysate

Bind RNA to column

Preparationof protein

RNA Isolation Protein Isolation


78776 50 preps78777 250 preps

Designed to remove dye-terminators from sequenc-ing reactions

�� 5 minute protocol�� Based on gel filtration technology�� Ideal for achieving excellent sequencing results�� Spin columns may be stored for long periods

The PrepEase Sequencing Dye Cleanup Kit is designed for easy and rapid separation of excess dye terminators from DNA sequencing reactions. The kit provides spin columns with a specialized gel filtration matrix which binds not only the excess dye terminators, but also impurities such as salts, traces of organic solvents, primers, and free nucleotides. The dye-free sequencing products are eluted from the column and are ready for analysis using standard methods.

Kit components:Includes PrepEase Dye Cleanup Columns and Col-lection Tubes.


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Acrylamide32800 25 gm 100 gm 500 gm 1 kg

bis-Acrylamide75821 25 gm 50 gm 100 gm 1 kg

RapidGel, 40% Liquid Acrylamide Stock Solution 75848 500 ml

Stop Solution 70724 1200 µl

TBE Buffer, 5X Solution 75891 1 L 5 L


PrepEase Sequencing Dye Cleanup Kits

Sequencing profile of plasmid DNA after treatment with the PrepEase Sequencing Dye Cleanup Kit.

Purification | Sequencing Dye Cleanup


Purification | Histidine-Tagged Protein Purification Kits

PrepEase Histidine-Tagged Protein Purification Mini Kit - High SpecificityBinding capacity up to 400 µgCulture volume up to 25 ml78791 50 preps

PrepEase Histidine-Tagged Protein Purification Midi Kit - High SpecificityBinding capacity up to 2.5 mgCulture volume up to 150 ml78793 50 preps

PrepEase Histidine-Tagged Protein Purification Maxi Kit - High SpecificityBinding capacity up to 5.0 mgCulture volume up to 300 ml78795 25 preps

Designed for fast, convenient, and high specificity purification of Histidine-tagged proteins

�� High specificity - due to single binding site to Histidine-tagged protein�� Low metal leaching - 5 binding sites to Ni2+

�� High purity > 95%

The PrepEase Protein Purification Kits for High Specificity are designed to purify recombinant polyhistidine-tagged (Histidine-tagged) recombinant proteins expressed in E. coli using immobilized metal ion affinity chromatography (IMAC). The kits may also be used to purify Histidine-tagged proteins from other expression systems such as insect cells, mammalian cells, and yeast. Purification may take place under standard, native conditions, or under denaturing conditions, depending on the solubility of the expressed protein.

PrepEase Protein Purification Kits are based on the use of novel, dry silica-based resin, precharged with Ni2+ ions. The High Specificity Kits utilize TED (tris-carboxymethyl ethylene diamine), a special chelating group, which enables strong and efficient binding of target proteins to the resin. TED is a strong pentadentate metal chelator, which occupies five of the six binding sites in the coordination sphere of the Ni2+ ion. The single remaining binding site can be exchanged with the histidine residues of the recombinant protein (Fig. 1).

Purification methods that utilize other chelators, such as NTA (nitrilotriacetic acid), may lead to metal leaching and/or non-specific binding of untagged proteins to the matrix. NTA chelating groups occupy four of the six binding sites in the coordination sphere of the Ni2+ ion, leaving two sites for protein binding. The additional chelating site of TED mini-mizes metal leaching during purification and reduces non-specific binding of untagged proteins to the ma-trix. This results in high specificity of Histidine-tagged protein binding to the resin, leading to higher purity of the eluted target protein (Fig. 2).

Kit components:Includes PrepEase High Specificity Histidine-tagged Protein Columns and all the necessary reagents for easy protein purification.


PrepEase Histidine-Tagged Protein Purification Kits - High Specificity

PrepEase Protein Purification Kits�� Novel – Based on dry silica based resin�� Less non-specific binding of contaminating proteins compared to Ni agarose beads�� Easy to elute protein off column�� Utilizes metal (Ni) affinity gravity-flow chromatography�� High stability against chelating and reducing agents�� Room temperature storage

Fig. 1. A: Silica bead bearing Ni2+ bound to TED, pentadentate metal chelator B: One histidine residue of the Histidine-tagged protein binding to the metal ion.

Binding of Histidine-Tagged Protein to PrepEase Ni-TED Resin

KDa

94

67

43

30

20.114.4

M CL F Wash Elution

10 20 50 100 200 250 mM Imidazole

PrepEase Ni-TED

KDa

94

67

43

30

20.114.4

M CL F Wash Elution

10 20 50 100 200 250 mM Imidazole

Ni-NTA AgarosePrepEase Silica-Based Resin Compared to Agarose Beads

Fig. 2. PrepEase High Specificity Ni-TED Resin compared to Ni-NTA Agarose.


Purification | Histidine-Tagged Protein Purification Kits

PrepEase Histidine-Tagged Protein Purification Mini Kit - High YieldBinding capacity up to 800 μgCulture volume up to 30 ml78801 50 preps

PrepEase Histidine-Tagged Protein Purification Midi Kit - High YieldBinding capacity up to 5 mgCulture volume up to 200 ml78803 50 preps

PrepEase Histidine-Tagged Protein Purification Maxi Kit - High YieldBinding capacity up to 10 mgCulture volume up to 375 ml78805 25 preps

Designed for fast, convenient, and high yield purifi-cation of Histidine-tagged proteins

�� High binding capacity - due to three-binding sites to Histidine-tagged proteins�� 2 times higher binding capacity than PrepEase High Specificity Purification Resin

The PrepEase Protein Purification Kits for High Specificity are designed to purify recombinant polyhistidine-tagged (Histidine-tagged) recombinant proteins expressed in E. coli using immobilized metal ion affinity chromatography (IMAC). The kits may also be used to purify Histidine-tagged proteins from other expression systems such as insect cells, mammalian cells, and yeast. Purification may take place under standard, native conditions, or under denaturing conditions, depending on the solubility of the expressed protein.

PrepEase Protein Purification Kits are based on the use of novel, dry silica-based resin, precharged with Ni2+ ions. Binding of protein is based on the interaction between the polyhistidine tag of the recombinant protein and immobilized Ni2+ ions. The High Yield Kits utilize IDA (iminodiacetic acid), a special chelating group, which enables strong and efficient binding of target proteins to the resin. IDA is a tri-dentate chelator which occupies three of the six binding sites in the coordination sphere of the Ni2+ ion. The remaining three coordination sites can be exchanged with histidine residues of the recombi-nant protein (Fig. 1).

In contrast to traditional IDA matrices, PrepEase Ni-IDA Resin contains an optimized, low density of IDA ligands that are created by a special manufacturing process. The non-saturating surface concentration of IDA eliminates non-specific interactions with contaminating proteins. As a result, PrepEase Ni-IDA Resin allows for higher target protein purity (Fig. 2).

Kit components:Includes PrepEase High Yield Histidine-tagged Protein Columns and all the necessary reagents for easy protein purification


PrepEase Histidine-Tagged Protein Purification Kits - High Yield

Fig. 1. Structure of PrepEase High Yield Ni-IDA Resin complexed with Ni2+.

PrepEase High Yield Ni-IDA Resin

Fig. 2. PrepEase High Yield Ni-IDA Resin compared to Ni-IDA Agarose.

KDa

9467

43

30

20.1

14.4

M CL Wash Elution

10 20 50 100 250 500 mM Imidazole

PrepEase Ni-IDA Ni-IDA Agarose

KDa9467

43

30

20.1

14.4

M CL Wash Elution

10 20 50 100 250 500 mM Imidazole

PrepEase Silica-Based Resin compared to Agarose Beads

PrepEase Protein Purification Kits�� Novel – Based on dry silica based resin�� Less non-specific binding of contaminating proteins compared to Ni agarose beads�� Easy to elute protein off column�� Utilizes metal (Ni) affinity gravity-flow chromatography�� High stability against chelating and reducing agents�� Room temperature storage


Purification | Histidine-Tagged & GST-Tagged Protein Purification

78796 30 gm

Binding capacity up to 10 mg protein per gm of resin

PrepEase Histidine-tagged High Specificity Purifica-tion Resin is a dry, silica-based resin, precharged with Ni2+ ions. The resin matrix, TED (Tris-carboxy-methyl ethylene diamine), is a pentadentate metal chelator which enables strong and efficient binding of target proteins. Metal leaching and non-specific binding of non-histidine proteins are significantly reduced, resulting in specific, high purity protein.


PrepEase Histidine-Tagged High Specificity Purification Resin

78806 30 gm 120 gm

Binding capacity up to 20 mg protein per gm of resin

PrepEase Histidine-tagged High Yield Purification Resin is a dry, silica-based resin, precharged with Ni2+ ions. The resin matrix, IDA (iminodiacetic acid), is a tridentate metal chelator which enables strong and efficient binding of target proteins. The PrepEase Ni-IDA Resin has multiple binding sites to Histidine-tagged protein, resulting in higher yields.


PrepEase Histidine-Tagged High Yield Purification Resin

78820 10 ml 100 ml

Designed for high quality purification of GST-tagged proteins by batch purification, gravity flow, or FPLC™.

�� Yields > 8 mg per 1 ml bed volume�� 60% higher binding capacity compared to Glutathione Sepharose™ 4B�� Flexible – perform batch purification, gravity flow or FPLC�� Easy regeneration – purify more protein over the life of the resin

PrepEase Protein Purification Glutathione Agarose 4B is a medium for single-step purification of glutathione-S-transferase (GST) fusion proteins by affinity chromatography. GST-tagged proteins are purified from cell lysates by their affinity to glutathione which is immobilized on agarose beads. Elution is performed with free glutathione in a non-denaturing, neutral pH buffer. PrepEase Protein Purification Glutathione Agarose 4B is appropriate for purifying GST-tagged proteins by gravity flow chromatography, FPLC, and batch processing.

Kit components:Includes PrepEase Protein Purification Glutathione Agarose 4B for single-step purification of glutathione-S-transferase (GST) fusion proteins by affinity chromatography.

Shipping and storage: Ships ambient. Store at 4°C.

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PrepEase Protein Purification Glutathione Agarose 4B

RNA AnalysismRNA Assay

RNA Transcription

RNA – Reverse Transcription

RNases

RNase Inhibitors

RNA Marker


mRNA AssayPoly(A) Tail-Length Assay Kit 158

RNA TranscriptionE. coli RNA Polymerase Holoenzyme 159Poly(A) Polymerase, Yeast 159T3 RNA Polymerase 160T7 RNA Polymerase 161

RNA - Reverse TranscriptionAMV Reverse Transcriptase 162M-MLV Reverse Transcriptase 162

RNases 163

RNase InhibitorsRNase Inhibitor (Human Placenta) 164RNase Inhibitor (Recombinant) 164

RNA MarkerLow Molecular Weight Marker, 10 - 100 nt 165

RNA Analysis Related Products 166

RNA Analysis | Table of Contents


76455 1 kit

Polyadenylation of eukaryotic mRNAs plays a critical role in RNA metabolism including stability, enhanced translation, nuclear export and miRNA mediated gene regulation. The Poly(A) Tail-Length Assay Kit allows you to quickly measure the poly(A) tails of multiple mRNAs from total RNA in only three hours. This kit utilizes a novel G/I extension technique which preserves the 3’ end of RNAs and ensures accurate measurement of poly(A) tail-length. And, all reaction components have been optimized so you get quality data quickly.

�� Measure the poly(A) tail of multiple mRNAs in under 3 hrs using common lab equipment �� Accurate measurement of poly(A) tail length compared to other assays (e.g. northern blotting)

� One kit includes optimized reagents for 5 G/I Tailing, 20 RT and 80 PCR reactions

The Poly(A) Tail-Length Assay Kit uses four key steps to enable poly(A) tail-length determination (Fig. 1).

In Step 1, poly(A) polymerase adds a limited number of guanosine and inosine residues to the 3’-ends of poly(A)-containing RNAs.

In Step 2, the tailed-RNAs are converted to DNA through reverse transcription using the newly added G/I tails as the priming sites.

In Step 3, PCR amplification products are generated using two primer sets. A gene-specific forward and reverse primer set designed upstream of the polyad-enylation site (e.g. the 3’-UTR) is produced as a con-trol for specific detection of the gene-of-interest. The second set of primers uses the gene-specific forward primer and the universal reverse primer provided with the kit to generate a product that includes the poly(A) tails of the gene-of-interest.

Finally, in Step 4, the PCR products are separated on an agarose or polyacrylamide gel (Fig. 2).

Kit components:Kit contains all necessary reagents for 5 G/I Tailing, 20 reverse transcription, and 80 PCR reactions.


References:1. Parker, R. and Song, H. (2004) Nat. Struct. Mol.

Biol. 11 121-127.2. Brodersen, D. E. et al. (2008) Biochim. Biophys.

Acta. 1779, 532-537.

3. Wu, L., Fan J. and Belasco, J. G. (2006) Proc. Natl. Acad. Sci. USA 103, 4034-4039.

4. Izaurralde, E. et al. (2009) RNA 15, 21-32.5. Martin, G. and Keller, W. (1998) RNA 4, 226-230.6. Gauss-Müller, V. et al. (2001) Nucleic Acids Res.

29 E57-7.

Poly(A) Tail-Length Assay Kit

Fig. 2. Comparison of human actin poly(A) tail-length in brain, muscle, liver and HeLa cell. One microgram total RNA and 4 μl diluted RT samples were used in G/I Tailing and PCR reactions, respectively. One half of each PCR reaction (12.5 μl) was analyzed by SYBR Gold-stained 6% non-denaturing polyacrylamide-TBE gel (A), and ethidium bromide-stained 2.5% agarose-TAE gel (B). With RT (+); No RT Negative Control (-); poly(A) tail PCR (A); gene-specific PCR (S); and marker (M).

Fig. 1. Outline of Poly(A) Tail-Length Assay.

RNA Analysis | mRNA Assay



Source:E. coli RNA Polymerase Holoenzyme is isolated from the rifampicin-sensitive strain BL21 and is sigma saturated.

Description:E. coli RNA Polymerase is a holoenzyme used for transcription of DNA and the production of labeled RNA for hybridization probes. The polymerase con-sists of 5 subunits designated α, α, β', β and σ. The enzyme is 100% saturated with sigma factor which is required for correct initiation of RNA synthesis on DNA templates containing bacterial and phage promoters.

Application:�� Synthesis of transcripts when it is not possible to transcribe DNA from a vector containing a phage RNA promoter

Properties:Molecular Weight: Approximately 450 kDa. The molecular weight for the subunits are 36.5 (each), 156, 151 and 70 kDa for α, β', β and σ subunits respectively.

Purity:Tested for contaminating non-specific exonucleases, endonucleases and ribonucleases.

Storage buffer:50 mM Tris-HCl (pH 7.5), 250 mM NaCl, 0.1 mM EDTA, 1.0 mM DTT, 50% glycerol.

Assay conditions:The reaction mixture (250 µl) contains 40 mM Tris-HCI (pH 7.5), 150 mM KCI, 10 mM MgCl2, 0.01% Triton X-100, 10 mM DTT, 0.02 µg/µl DNA template, 0.5 mM of each rNTP, and enzyme. Incubation is at 37°C for 10 minutes.

Unit definition:One unit is the amount of enzyme required to cata-lyze the incorporation of 1 nmol of a ribonucleoside triphosphate into RNA in 10 minutes at 37°C.

Concentration:1 unit/µl


References:1. Burgess, R. R. and Travers, A. A. (1970) Fed Proc.

29, 1164-1169.2. Burgess, R. R., Travers, A. A., Dunn, J. J. and Bautz,

E. K. (1969) Nature 221, 43-46.3. Fujiki, H., Palm, P., Zillig, W., Calendar, R. and

Sunshine, M. (1976) Mol. Gen. Genet. 145, 19-22.

E. coli RNA Polymerase Holoenzyme (RNA Nucleotidyl Transferase, E.C.2.7.7.6)


Poly(A) Polymerase Reaction Buffer, 5X(Included with each pack size of enzyme)74226 1 ml

Source:E. coli strain containing an overproducing clone of Yeast Poly(A) Polymerase.

Description:Poly(A) Polymerase catalyzes the addition of adeno-sine residues onto the 3'-ends of RNA(1,2). It can be used to add poly(A) tails to RNA in the first step of cloning(3). The reaction requires Mn2+ or Mg2+, ATP as substrate and any RNA containing 3' hydroxyl ter-mini as primer. Longer RNA molecules are somewhat better primers than short oligomers(4). Substitution of cordycepin-5'-triphosphate (3'-dATP) for ATP results in addition of a single 3'-dA residue to the ends of the RNA, a useful technique for labeling RNA at the 3'-end(5)*.

In comparative studies, Yeast Poly(A) Polymerase works more efficiently than E. coli Poly(A) Polym-erase for RNA oligonucleotide-labeling and poly(A) tailing. Shorter incubation times are required for the yeast enzyme and it is found to label both long and short substrates equally well. Poly(A) Polymerase is recommended over T4 RNA Ligase for 3'-end label-ing of long RNA molecules.

*Note: Various modified nucleotides can also be used to label the 3' end of RNA using Yeast Poly(A) Polymerase(5).

Applications:�� Addition of poly(A) tails to RNA�� Labeling the 3' ends of RNA

Purity:Ribonuclease free.

Storage buffer:20 mM Tris-HCl (pH 8.0), 50 mM KCl, 0.5 mM DTT, 50% glycerol.

Assay conditions:25 mM Tris-HCl (pH 7.0), 40 mM KCl, 0.5 mM MnCl2, 0.5 mM EDTA, 0.5 mM DTT, 0.2 mg/ml BSA, 10% glycerol, 3.3 µM radiolabeled ATP, 0.5 mM ATP, 6.5 µg poly(A) (~100 bases), poly(A) polymerase. After incubation at 37°C for 10 minutes, acid insoluble radioactivity is determined.

Unit definition:One unit is the amount of enzyme which incorpo-rates 1 pmol AMP into acid-insoluble material at 37°C in 1 minute.


Functional test:3'-end labeling of a ribonucleotide with cordycepin-5'-triphosphate.

Functionally tested 5X Poly(A) Polymerase Reaction Buffer (1 ml included, PN 74226):100 mM Tris-HCl (pH 7.0), 3.0 mM MnCl2, 0.1 mM EDTA, 1 mM DTT, 500 µg/ml acetylated BSA, 50% glycerol.


References:1. Sippel, A. E. (1973) Eur. J. Biochem. 37, 31-40.2. Edmonds, M. (1982) in The Enzymes, 3rd edition,

ed. P. D. Boyer (Academic Press, New York) 15, 217-244.

3. Gething, M. J., Bye, J., Skehel, J. and Waterfield, M. (1980) Nature 287, 301-306.

4. Sano, H. and Feix, G. (1976) Eur. J. Biochem. 71, 577-583.

5. Martin, G. and Keller, W. (1998) RNA 4, 226-230.

Related products


Poly(A) Polymerase, Yeast

RNA Analysis | RNA Transcription


Standard concentration, 20 units/µl70051Y 1,000 units70051Z 5,000 unitsHigh concentration, 200 units/µl70204 1,000 units 5,000 units

Source:E. coli strain containing an overproducing clone of T3 RNA Polymerase.

Description:T3 RNA Polymerase has a high specificity for bacteriophage T3 promoter sequences. It is used in the synthesis of large amounts of specific RNA transcripts from vectors containing the T3 promoter. Transcripts may be used as probes in Northern and Southern blots(1), substrates for in vitro translation studies(2), substrates for RNA processing studies(3) and for exon and intron mapping of genomic DNA(1). The enzyme may be used with both radioactively la-beled(4) and hapten-labeled (fluorescein, digoxigenin, biotin, etc.) nucleoside triphosphates(5). T3 RNA Polymerase is also used to generate capped mRNA for expression studies in oocytes and other cells(6).

Applications:�� Production of RNA transcripts�� Production of capped or modified RNA transcripts

Purity:Greater than 90% pure as determined by SDS-PAGE. Tested for contaminating non-specific endonucle-ases, exonucleases and ribonucleases.

Storage buffer:20 mM HPO4 (pH 7.7), 100 mM NaCl, 1 mM EDTA, 10 mM DTT, 1.0% CHAPS, 50% glycerol.

Assay conditions:The reaction mixture contains 1X Transcription Buffer, 0.5 mM each NTP, 1 µCi labeled CTP, 1 µg of T3 promoter containing DNA and T3 RNA Polymerase. Incubation is at 37°C for 15 minutes (50 µl reaction volume).

Unit definition:One unit catalyzes the incorporation of 1 nmol of CMP into polynucleotide fraction bound to a positively charged DE-81 filter paper in 60 minutes at 37°C using a linearized 17 promoter containing 280 nt template.

Concentration:Standard concentration (PN 70051Y/Z): 20 units/µlHigh concentration (PN 70204): 200 units/µl

Functionally tested 10X T7/T3 RNA Polymerase Transcription Buffer:400 mM Tris-HCl (pH 8.0), 60 mM MgCl2, 100 mM DTT, 100 mM NaCl, 20 mM spermidine trihydro-chloride.


References:1. Melton, D. A., Krieg, P. A., Regagliati, M. R.,

Maniatis, T., Zinn, K. and Green, M. R. (1984) Nucl. Acids Res. 12, 7035-7056.

2. Krieg, P. A. and Melton, D. A. (1984) Nucl. Acids Res. 12, 7057-7070.

3. Green, M. R., Maniatis, T. and Melton, D. A. (1983) Cell 32, 681-694.

4. Davanloo, P., Rosenberg, A. H., Dunn, J. J. and Studier, F. W. (1984) Proc. Natl. Acad. Sci. USA 81, 2035-2039.

5. Langer, P. R., Waldrop, A. A. and Ward, D. C. (1981) Proc. Natl. Acad. Sci. USA 78, 6633-6637.

6. Sambrook, J. and Russell, D. W. (2001) Molecular Cloning: A Laboratory Manual, 3rd Edition, Cold Spring Harbor, New York, 9.87-9.88.

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Dithiothreitol (DTT), 0.1 M Solution70726 150 μl 1 ml 5 ml

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T7 RNA PolymeraseStandard Concentration, 20 units/μl70047Y 6,000 unitsHigh Concentration, > 200 units/μl70001Z 30,000 units


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NTPs, Set of Four, (ATP, CTP, GTP, UTP), 100 mM Solutions, 4 x 25 µmol77245 4 x 25 µmol

T3 RNA Polymerase (RNA Nucleotidyl Transferase, E.C.2.7.7.6)



Standard concentration, 20 units/µl70047Y 6,000 unitsHigh concentration, >200 units/µl70001Z 30,000 unitsCustoms by request

Source:E. coli strain containing an overproducing clone of T7 RNA Polymerase(1).

Description:T7 RNA Polymerase is a single-subunit enzyme produced by bacteriophage T7(2). It is highly specific for T7 promoter and terminator sequences(2,3). It has been widely used for the rapid synthesis in vitro of specific RNAs. These transcripts can be used directly as substrates for studies of RNA structure or me-tabolism. If the transcripts are suitably labeled, they can also be used as sensitive hybridization probes. T7 RNA Polymerase, along with chain-terminating nucleoside triphosphates(4), have also been used for the direct sequencing of DNA. T7 RNA Polymerase is also used to generate capped mRNA for expression studies in oocytes and other cells(5).

Applications:�� Production of RNA transcripts for hybridization probes�� Production of large amounts of discrete sized RNA�� Production of capped or modified RNA transcripts

Properties:Molecular Weight: 98.8 kDaOptimum pH: 7.7 - 8.3Optimum Temperature: 37°CRequirement for Divalent Cation: Mg2+ (optimal

concentration is 20 mM)Michaelis Constants(2): 40 µM ATP, 160 µM GTP,

60 µM UTP, 80 µM CTP

Purity:Greater than 99% pure as determined by SDS-PAGE. Tested for contaminating nonspecific endonucleases, exonucleases and ribonucleases.

Storage buffer:20 mM HPO4, pH 7.7, 100 mM NaCl, 1 mM EDTA, 10 mM DTT, 1% CHAPS and 50% glycerol.

Assay conditions:The reaction mixture contains 1X Transcription Buffer, 0.5 mM each NTP, 1 µCi labeled CTP, 1 µg of T7 promoter containing DNA and T7 RNA Polymerase. Incubation is at 37°C for 15 minutes (50 µl reaction volume).

Unit definition:One unit catalyzes the incorporation of 1 nmol of CMP into polynucleotide fraction bound to a positively charged DE-81 filter paper in 60 minutes at 37°C using a linearized T7 promoter containing 280 nt template.

Concentration:Standard concentration (PN 70047Y): 20 units/µlHigh concentration (PN 70001Z): >200 units/µl

Functional test:Transcription from plasmid containing T7 promoter; >10 µg of RNA can be produced from 1 µg of supercoiled template.

Functionally tested 10X T7/T3 RNA Polymerase Transcription Buffer:400 mM Tris-HCl, pH 8.0, 60 mM MgCl2, 100 mM DTT, 100 mM NaCl, 20 mM spermidine trihydro-chloride.


References:1. Tabor, S. and Richardson, C. C. (1985) Proc. Natl.

Acad. Sci. USA 82, 1074-1078.2. Chamberlin, M. and Ring J. (1973) J. Biol. Chem.

248, 2235-2244, 2245-2250.3. Chamberlin, M. and Ryan, T. (1982) in The

Enzymes, 3rd edition, ed. P. D. Boyer (Academic Press, New York.) 15, 87-108.

4. Axelrod, V. D. and Kramer, F. R. (1985) Biochemistry 24, 5716-5723.

5. Sambrook, J. and Russell, D. W. (2001) Molecular Cloning: A Laboratory Manual, 3rd Edition, Cold Spring Harbor, New York, 9.87-9.88.

6. Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A. and Struhl, K. (1998) Current Protocols in Molecular Biology (John Wiley and Sons, Inc.).

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NTPs, Set of Four, (ATP, CTP, GTP, UTP), 100 mM Solutions, 4 x 25 µmol77245 4 x 25 µmol

T7 RNA Polymerase (RNA Nucleotidyl Transferase, E.C.2.7.7.6)




Source:Purified virion of Avian myeloblastosis virus.

Description:AMV Reverse Transcriptase catalyzes the polymeriza-tion of DNA using template DNA, RNA or RNA:DNA hybrids(1). It consists of two polypeptide chains, one of which contains a 5'→3' polymerase activity and the other an RNase H activity. Reverse transcriptase requires Mg2+ or Mn2+ and is used for first strand synthesis of complementary DNA (cDNA) from mRNA template(2). Under proper conditions, high yields of full-length cDNA can be obtained with AMV Reverse Transcriptase(3). AMV Reverse Transcriptase is also useful in the amplification of RNA(4).

Applications:�� Synthesis of cDNA for PCR, cloning and hybridization probes�� Filling-in and labeling the 3' termini of DNA with 5' protruding ends�� RNA sequencing�� Amplification of RNA(4)

�� Primer extension assays

Properties:Molecular Weight: 157 kDaInactivation: 75°C for 10 minutes or by adding 2 µl

of 0.5 M EDTA for a 50 µl reaction.

Purity:Tested for contaminating endonucleases, exonucle-ases and ribonucleases.

Storage buffer:200 mM potassium phosphate (pH 7.2), 2 mM DTT, 0.2% Triton X-100, 50% glycerol.

Assay conditions:The reaction mixture (100 µl) contains 50 mM Tris-HCI (pH 8.3), 6 mM MgCl2, 40 mM KCI, 0.5 mM radiolabeled TTP, 0.4 mM poly(rA)n:oligo(dT)50. Incubation is at 37°C for 10 minutes.

Unit definition:One unit is the amount of enzyme required to incor-porate 1 nmol of radiolabeled nucleotide into acid insoluble product in 10 minutes at 37°C.


Functional test:RT-PCR amplification on generation of 0.5 kb and 1.5 kb RT-PCR products.

Functionally tested 5X AMV RT Reaction Buffer (1 ml included):250 mM Tris-HCl (pH 8.3), 40 mM MgCl2, 250 mM NaCl, 5 mM DTT

Shipping and storage:Shipped on dry ice. Store at -20°C. Avoid freeze-thaw cycles, which result in loss of enzyme activity.

References:1. Kacian, D. L. (1977) in Methods in Virology, eds.

K. Maramorosch and H. Koprowski 6, 143-184.2. Goodman, H. M. and MacDonald, R. J. (1979)

Methods Enzymol. 68, 75-90.3. Berger, S. L., Wallace, D. M., Puskas, R. S. and

Eschenfeldt, W. H. (1983) Biochemistry 22, 2365-2372.

4. PCR Protocols: A Guide to Methods and Applications (1990) eds. M. A. Innis, D. H. Gelfand, and John J. Sni, (Academic Press, New York), 23.

AMV Reverse Transcriptase

78306 25,000 units 100,000 unitsCustoms by request

M-MLV Reaction Buffer, 5X(Included with each pack size of enzyme)71505 1 ml

Source:E. coli strain containing an overproducing clone of M-MLV Reverse Transcriptase.

Description:M-MLV Reverse Transcriptase catalyzes the polymerization of DNA using template DNA, RNA or RNA:DNA hybrids(1). Full-length copies of large mRNAs, >10 kb, may be synthesized. M-MLV Reverse Transcriptase has a much lower RNase H activity than AMV Reverse Transcriptase resulting in high yields of full length cDNA(2). This makes M-MLV Reverse Transcriptase very useful in cDNA synthesis and RT-PCR.

For added convenience and optimal RT-PCR, try the USB RT-PCR Kits (PN 78350 and 78355) and RT-PCR Master Mixes (PN 71185 and 78370).

Applications:�� RT-PCR�� Synthesis of first strand cDNA for PCR, cloning and hybridization probes�� Filling-in and labeling the 3' termini of DNA with 5' protruding ends�� Amplification of RNA�� Primer extension assays

Properties:Molecular Weight: 71 kDa (monomeric).Inhibitors: Polyamines, phosphate, pyrophosphates

and lithium chloride(3,4).Inactivation: 75°C for 10 minutes or by adding 2 µl

of 0.5 M EDTA for a 50 µl reaction.

Purity:Greater than 90% pure as determined by SDS-PAGE. Tested for contaminating endonucleases, exonucle-ases and ribonucleases.

Storage buffer:20 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 0.1 mM EDTA, 1 mM DTT, 0.01% Igepal CA-630, 50% glycerol.

Assay conditions:The reaction mixture (25 µl) contains 50 mM Tris-HCI (pH 8.3), 40 mM KCl, 6 mM MgCl2, 1 mM DTT, 0.4 mM poly(rA)-oligo(dT)12 - 18, 0.5 mM radiolabeled TTP and 0.1 - 0.5 units enzyme. Incubation is at 37°C for 10 minutes.

Unit definition:One unit is that amount of enzyme required to incorporate 1 nmol of deoxynucleotide into DE-81 filter-binding material in 10 minutes at 37°C using poly (rA)-oligo(dT)12-18 as template-primer.


Functional test:RT-PCR amplification on generation of 0.5 kb and 1.5 kb RT-PCR products.

Functionally tested 5X M-MLV Reaction Buffer (1 ml included, PN 71505):250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2, 50 mM DTT.


References:1. Roth, M. J., Tanese, N. and Goff, S. P. (1985)

J. Biol. Chem. 260, 9326-9335.2. Sambrook, J. and Russell, D. W. (2001) Molecular

Cloning: A Laboratory Manual, 3rd Edition, Cold Spring Harbor, New York, 8.48.

3. Gerard, G. F. and D’Alessio, J. M. (1993) Methods In Molecular Biology 16, Humana Press, NJ, 73-93.

4. Sambrook, J. and Russell, D. W. (2001) Molecular Cloning: A Laboratory Manual, 3rd Edition, Cold Spring Harbor, New York, A8.16.

M-MLV Reverse Transcriptase (E.C.2.7.7.49)

RNA Analysis | RNA—Reverse Transcription


Prod. No. Product Description Size

27-0330-02 RNase I 1 gm See page 79 RNase I catalyzes the hydrolysis of 3'→5' phosphodiester bonds of RNA with the formation of oligoribonucleotides terminating in pyrimidine 2'→3' cyclic phosphate.

27-0323-01 RNase I ‘A’, Powder 100 mg See page 79 RNase I ‘A’ specifically cleaves single-stranded RNA at 3' phosphate linkages of pyrimidine residues leaving pyrimidine 3' phosphates and oligonucleotides with terminal pyrimidine 3' phosphates.

78020Y RNase A, Lyophilized 100 mg See page 79 RNase A specifically cleaves single-stranded RNA at 3' phosphate linkages of pyrimidine residues leaving pyrimidine 3' phosphates and oligonucleotides with terminal pyrimidine 3' phosphates.

70194Y/Z RNase A, Molecular Biology Grade Solution 1 mg See page 80 RNase A specifically cleaves single-stranded RNA at 3' 5 mg phosphate linkages of pyrimidine residues leaving pyrimidine 3' phosphates and oligonucleotides with terminal pyrimidine 3' phosphates.

70054Y/Z RNase H 250 units See page 80 E. coli RNase H is an endoribonuclease that degrades the 1,000 units RNA portion of DNA:RNA hybrids.

RNases

RNA Analysis | RNases

Customized solutions

�� Our experts in custom and OEM reagents will work with you at your site and from our headquarters�� Our over 3,000 catalog products can be tailored to meet your specific needs�� Custom fill sizes, concentrations, and formulations�� Volume fills from microliter to liters�� Weight fills of milligrams to kilograms�� Customer-specific quality control assays�� Liquid conversion to powder in pre-weighed packaging

Most of our products can be tailored to meet your specifications. With our high standards, ISO certification, and scalable purchasing, we are well positioned to offer you a wide range of customized quality products.




Source:Human placenta

Description:Human Placental RNase Inhibitor (HPRI) binds and inhibits a broad range of eukaryotic RNases, includ-ing RNase A, RNase B, RNase C and human placen-tal RNase. HPRI is purified from human placenta by affinity chromatography on an immobilized RNase A column. It forms a 1:1 complex with RNase A and inhibits RNase activity non-competitively (Ki = 3 x 1010 M). It can be added directly to reaction mixtures containing RNA. HPRI differs from other competitive inhibitors in that it can be easily removed from reac-tion mixtures by phenol extraction.

RNase Inhibitor is not effective against RNase 1, RNase T1, S1 Nuclease, RNase H or RNase from Aspergillus. To maintain activity, HPRI requires a minimum of 1 mM DTT and is active over a broad pH range (pH 5.5-9.0).

Applications:�� cDNA synthesis(2)

�� In vitro translation(3)

�� In vitro transcription(4)

�� Polysome isolation�� RT-PCR�� Isolation of mammalian cell samples that contain a mRNA-protein complex(3)

�� Separation and identification of specific ribonuclease activities(5)

Properties:Activators: Requires DTT for activityInhibitors: RNases will recover activity after

inactivation at temperatures greater than 55°C or in 7 M urea.

Purity:Tested for contaminating non-specific endonu-cleases, exonucleases, ribonucleases and latent ribonucleases.

Storage buffer:20 mM HEPES-KOH (pH 7.6), 50 mM KCl, 8 mM DTT, 50% glycerol.

Unit definition:One unit inhibits the activity of 5 ng of RNase A by 50%(1). This inhibitor activity is determined by its ability to inhibit the hydrolysis of cyclic 2' 3'-CMP by RNase A.

Concentration:≥ 20 units/µl

Shipping and storage:Shipped on dry ice. Store at -20°C. Avoid repeated freeze/thaw cycles.

References:1. Blackburn, P. (1979) J. Biol. Chem. 254, 12484-

12487.2. de Martynoff, G., Pays, E. and Vassart, G. (1980)

Biochem. Biophys. Res. Commun. 93, 645-653.3. Scheele, G. and Blackburn, P. (1979) Proc. Natl.

Acad. Sci. USA 76, 1898-1902.4. Nielson, D. A. and Shapiro, D. J. (1986) Nucl. Acids

Res. 14, 5936.5. Eichler, D. C., Tatar, T. F. and Lasater, L. S. (1981)

Biochem. Biophys. Res. Commun. 101, 396-403.

RNase Inhibitor (Human Placenta)


Source:E. coli strain containing an overproducing clone of human placenta ribonuclease inhibitor.

Description:Recombinant Human Placental RNase Inhibitor (rHPRI) binds and inhibits a broad range of eukary-otic RNases, including RNase A, RNase B, RNase C and human placental RNase. HPRI forms a 1:1 complex with RNase A and inhibits RNase activity non-competitively (Ki = 3 x 1010 M). It can be added directly to reaction mixtures containing RNA. HPRI differs from other competitive inhibitors in that it can be easily removed from reaction mixtures by phenol extraction.

RNase Inhibitor is not effective against RNase 1, RNase T1, S1 Nuclease, RNase H or RNase from Aspergillus. To maintain activity, HPRI requires a minimum of 1 mM DTT and is active over a broad pH range (pH 5.5-9.0).

Applications:�� cDNA synthesis(2)

�� In vitro translation(3)

�� In vitro transcription(4)

�� Polysome isolation�� RT-PCR�� Isolation of mammalian cell samples that contain a mRNA-protein complex(3)

�� Separation and identification of specific ribonuclease activities(5)

Properties:Molecular Weight: 50 kDaIsoelectric Point: 4.7Activators: DTTInhibitors: RNases will recover activity after

inactivation at temperatures greater than 55°C or in 7 M urea.

Purity:Greater than 90% pure as determined by SDS-PAGE. Tested for contaminating non-specific endonucle-ases, exonucleases and ribonucleases.

Storage buffer:20 mM HEPES-KOH (pH 7.6), 50 mM KCl, 8 mM DTT, 50% glycerol.

Unit definition:One unit inhibits the activity of 5 ng of RNase A by 50%(1).


Shipping and storage:Shipped on dry ice. Store at -20°C. Avoid repeated freeze/thaw cycles.

References:1. Blackburn, P. (1979) J. Biol. Chem. 254, 12484-

12487.2. de Martynoff, G., Pays, E. and Vassart, G. (1980)

Biochem. Biophys. Res. Commun. 93, 645-653.3. Scheele, G. and Blackburn, P. (1979) Proc. Natl.

Acad. Sci. USA 76, 1898-1902.4. Nielson, D. A. and Shapiro, D. J. (1986) Nucl. Acids

Res. 14, 5936.5. Eichler, D. C., Tatar, T. F. and Lasater, L. S. (1981)

Biochem. Biophys. Res. Commun. 101, 396-403.

RNase Inhibitor (Recombinant)

RNA Analysis | RNase Inhibitors


76410 100 µl

Description:The Low Molecular Weight Marker is a set of 14 bands between 10 - 100 nucleotides, designed for size determination of single-stranded oligonucleotide fragments.

The marker is suitable for use in either polyacryla-mide or agarose gel electrophoresis. The recom-mended concentration for agarose is 2.5% and for polyacrylamide is 12 - 15%. This marker should be diluted with TE Buffer and the supplied Gel Loading Buffer to the desired loading concentration.

The Low Molecular Weight Marker can be used to generate a 5'-end radiolabeled marker for size comparison of small, single-stranded oligonucleotide fragments such as miRNA, siRNA, tRNA, snRNA, snoRNA, or ssDNA. The Low Molecular Weight Marker is also useful when the molecule of interest is less than 100 nucleotides in length. This marker is not designed for precise quantification of nucleic acid mass and should only be used as a size marker.

2X Formamide Loading Buffer and 6X Glycerol Load-ing Buffer are included for convenience to prepare both the marker and samples for gel electrophoresis.

Fragment sizes: 100 nt 40 nt 90 nt 35 nt 80 nt 30 nt 70 nt 25 nt 60 nt 20 nt 50 nt 15 nt 45 nt 10 nt

Functional test: Meets or exceeds criteria for 5'-end labeling with isotope and polyacrylamide gel electrophoresis.

Components:Low Molecular Weight Marker, 100 µl (50 ng/µl each

fragment)2X Formamide Loading Buffer6X Glycerol Loading Buffer

2X Formamide Loading Buffer (1 ml included): Consists of bromophenol blue, xylene cyanol FF, EDTA and formamide.

6X Glycerol Loading Buffer (1 ml included): Consists of orange G, xylene cyanol FF and glycerol.

Shipping and storage:Shipped on ice. Store at 4°C or at -20°C for long term storage.

To use:1. Mix 2 µl (100 ng each fragment) of Marker with

2 µl of 2X Formamide Loading Buffer, heat at 95°C for 3 minutes, quick cool on ice, and load the entire volume per lane on polyacrylamide gel.

2. Mix 5 µl (250 ng each fragment) of Marker with 1 µl of 6X Glycerol Loading Buffer and load the entire volume per lane on agarose gel.

3. Visualize with dyes for detecting single-stranded oligonucleotide fragments.

4. For 5'-end labeling reaction, use 1 µl (50 ng each fragment) of Marker with OptiKinase and [γ-32P]-ATP or [γ-33P]-ATP.

Related products – miRNA Detection


Related products – UItrapure electrophoresis reagents

Ammonium Persulfate 76322 100 gm



TAE Buffer, 10X Solution 75904 1 L 5 L


TE Buffer, 1X Solution75893 10 x 1 ml 100 ml 500 ml

TEMED76320 100 ml 100 gm 500 gm

Low Molecular Weight Marker, 10 – 100 nt

Fig. 1. Low Molecular Weight Marker on 12-15% UREA-polyacrylamide gel. A. 50 ng of SYBR® Gold stained Marker. B. 0.5 ng of 5' end labeled Marker with [γ-33P]-ATP and OptiKinaseTM. C. 0.05 ng of 5' end labeled Marker with [γ-32P]-ATP and OptiKinase.

100908070

60

5045

40

35

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10

100908070

60

5045

40

35

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10090807060

5045

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A B Cnt nt nt

RNA Analysis | RNA Marker



76410 Low Molecular Weight Marker, 10-100 nt 100 µl

RNA Marker


77241 ATP 100 mM 25 μmol (250 μl) Adenosine-5'-Triphosphate Ultrapure

77243 GTP 100 mM 25 μmol (250 μl) Guanosine-5'-Triphosphate Ultrapure

77245 NTPs, Set of Four (ATP, CTP, GTP, UTP) 100 mM 4 x 25 μmol (250 μl) Nucleoside Triphosphates 4 NTPs per pack (pk) Ultrapure 1 pk

Related Products

Oligo p(dT)


19817 Oligo p(dT)12-18 Sodium Salt 5 units 25 units 100 units

RNA Electrophoresis


76322 Ammonium Persulfate (APS) 10 gm 100 gm 1 kg

75827 Glycerol Tolerant Gel (GTG) Buffer, 20X Solution 1 L

75848 RapidGel, 40% Liquid Acrylamide Stock Solution 500 ml

75891 TBE Buffer, 5X Solution 1 L 5 L

76320 TEMED 100 ml 100 gm 500 gm

75826 Urea 1 kg 5 kg

Ribonucleotides (NTPs), Sodium Salt Solutions

RNA Analysis | Related Products


Related Products continued…

RNA Ligation


21245 T4 RNA Ligase 600 units

RNA Purification


78766 PrepEase RNA Spin Kit 50 preps

78878 PrepEase mRNA MiniSpin Kit 12 preps

RNAse-Free Buffers & Solutions


75901 Ammonium Acetate (NH4OAc), 5 M Solution 100 ml 500 ml

15694 EDTA, 0.5 M Solution 100 ml 500 ml

78641 Magnesium Chloride (MgCl2), 1 M Solution 10 x 1 ml 100 ml

75896 Potassium Chloride (KCl), 2 M Solution 100 ml

75888 Sodium Chloride (NaCl), 5 M Solution 100 ml 500 ml 1 L

22637 Tris/HCl, 1 M Solution, pH 7.0 100 ml 500 ml 1 L

22638 Tris/HCl, 1 M Solution, pH 8.0 100 ml 500 ml 1 L

71786 Water, Nuclease-Free 10 x 1 ml 100 ml 500 ml 1 L 5 L

70783 Water, RNase-Free, DEPC Treated 25 ml 10 x 1 ml 100 ml 500 ml 1 L

RNA Analysis | Related Products

DNA SequencingNGS Library Prep

Thermo Sequenase™ Kits

Sequenase Enzyme & Kits

Sequencing Primers

Sequencing Tips


DNA Sequencing | Table of Contents

Next Generation Sequencing (NGS) Library PrepPrep2Seq™ DNA Library Prep Kit for

Illumina® platform 170Prep2Seq Multiplex Oligo Adapters for

Illumina® platform 171

Thermo Sequenase KitsThermo Sequenase Cycle Sequencing Kit 172Thermo Sequenase Dye Primer Manual Cycle

Sequencing Kit 173

Sequenase Enzyme & KitsSequenase Version 2.0 DNA Polymerase 174Sequenase Version 2.0 DNA Sequencing Kit 175Sequenase PCR Product Sequencing Kit 176Sequenase Quick Denature Plasmid DNA

Sequencing Kit 177

Sequencing PrimersPrimers for DNA Sequencing 178

Sequencing TipsDetermining Primer Concentration 178DNA Sequencing Gel Formulations 178Use of Terminal Deoxynucleotidyl Transferase

(TdT) to Resolve Bands Across Four Lanes (BAFLs) 179

Relief from Compression Artifacts in DNA Sequencing Using Formamide Gels 180

Prevent Weak Sequencing Band Intensities by Using Pyrophosphatase 181



Generate high quality next generation sequencing (NGS) libraries suitable for sequencing on Illumina® platforms in less time with Prep2Seq DNA Library Prep Kit.

DNA library prep is critical to the precision and accuracy of NGS. The Prep2Seq DNA Library Prep Kit comes with all of the reagents necessary for end repair, A-tailing, and ligation, and can be completed in less than two hours.

Our kit is fast, efficient, and optimized. It efficiently converts 3’ and 5’ overhangs into 3’ A-overhang ends for ligation to T-tailed DNA adapters and reduces the potential for bias with our superior A-tailing and ligation steps. We offer significant improvement in the quality of sequence-ready libraries by eliminating chimeras and reducing side reaction fragments.

The Prep2Seq DNA Library Prep Kit combines end re-pair and A-tailing steps, decreasing the total number of steps to achieve quality NGS library preparation. Only one cleanup step is required to attain peak performance on the Illumina® sequencing platform and only 100 ng of input is needed.

Full kit offering�� Single kit that includes all reagents for sample prep in a slimmed down protocol.�� 24 bar code adapter sequences compatible with Illumina® technology are sold separately:

Prep2SeqMultiplexOligoAdaptersforIllumina - Index Adapter Set I–adapter sequences 1-12 - Index Adapter Set II–adapter sequences 13-27

Time savings with a more efficient method for NGS sample prep�� Improved A-tailing reaction with proprietary enzymes, cloning know-how, and optimized buffers.�� By combining the end repair and A-tailing steps, only one cleanup step is needed compared to the two or three steps in most other kits (saving up to two and a half hours in the lab).

The Prep2Seq DNA Library Prep Kit offers the highest adapter ligation efficiency available. By combining end repair and A-tailing, the Prep2Seq DNALibrary Prep Kit is able to reduce total NGS library prep time to 110 minutes and limit bead cleanup to a single step.

Rigorous comparison studies to competitor kits show that the Prep2Seq DNA Library Prep Kit generates impressive sequencing data.

Prep2Seq™ DNA Library Prep Kit for Illumina® platform

DNA Sequencing | Next Generation Sequencing Library Prep

Illumina® TruSeq® Nano

140 min

NEBNext® Ultra™

65 min

USB Prep2Seq™

40 minHands-on Lab Time

Walk-Away Time

USB - 110 total min

NEB - 155 total min

Illumina - 210 total minIllumina® TruSeq® Nano

70 min

NEBNext® Ultra™

90 min

USB Prep2Seq™

70 min

Table 1. Validated performance of the Prep2Seq DNA Library Prep Kit compared to competition.


Total Reads 170,507,429 171,139,925 176,073,965

Mapped reads to a single locus 142,223,440 143,567,898 147,688,157

% Mapped reads to a single locus 83.41% 83.89% 83.88%

Mapped reads to multiple loci 12,270,818 11,794,099 12,266,301

% Mapped reads to multiple loci 7.20% 6.89% 6.97%

Unmapped reads 16,013,171 15,777,928 16,119,507

% Unmapped reads 9.39% 9.22% 9.15%

The HiSeq® 2500 raw reads were aligned to the human genome hg19 release using Bowtie, allowing 2 mismatches for the entire read length. Mapped reads aligning to a single locus, mapped reads aligning to multiple loci, as well as unmapped reads from the 3 libraries are summarized in Table 1.

Table 2. GC content present in mapped and unmapped reads for the 3 library prep methods.


% GC: Mapped reads to a single locus 42.27% 40.01% 41.10%

% GC: Mapped reads to multiple loci 43.53% 41.65% 42.55%

% GC: Unmapped reads 40.90% 38.79% 40.64%

In order to determine if the end-repair and adaptor ligation may have contributed to some library bias we compared the overall GC content in mapped and unmapped reads for each of the 3 kits. As shown in Table 2, no substantial difference in GC content was introduced during the preparation methods of the 3 kits.

Time Comparison


min Ligation set up

NEB Next Ultra

USB Prep2Seq 5

min Ligation set up

30min Incubation

40min Incubation


Incubation55min

Clean up

5min

End repair set up

30min Incubation

35min

Clean up

5 30min Incubation

5

5 10min Incubation

90min Clean up

30min Clean up


min A-tail set up





min Ligation set up



DNA Sequencing | Next Generation Sequencing Library Prep

Prep2Seq™ DNA Library Prep Kit for Illumina® platform continued...

79800 1 KT Set 1(Contains adapter sequences 1-12)

2 KT Set 2(Contains adapter sequences 13-27)

Prep2Seq Multiplex Oligo Adapters allow sample multiplexing in NGS reactions. These are HPLC purified adapter duplex sets that are bar coded and designed for multiplex sample preparation for Next Generation Sequencing on Illumina® platforms.


adapters 1-12.20 reactions per adapter Each adapter vial contains 4.8 nmol, 80 μl.


adapters 13-16, 18-23, 25, and 27. 20 reactions per adapterEach adapter vial contains 4.8 nmol, 80 μl.

Prep2Seq Multiplex Oligo Adapters for Illumina® platform

The Prep2Seq DNA Library Prep Kit ensures efficiency of ligation to T-tailed adapters by optimizing the A-tailing step. This enzymology focus on A-tailing ensures that end bias is reduced, and the formation of blunt- and G-tailed ends is minimized. Most enzymes used for A-tailing are inefficient and result in a large amount of blunt-end DNA that can ligate as concatemers, forming chimeric false joints and compromising sequencing data. Effective A-tailing is crucial in preparing efficient DNA adapter libraries that are ready for sequencing.

Also available separately are barcoded adapter sets that enable multiplexing of up to 24 samples. Prep2Seq Multiplex Oligo adapters for Illumina® systems are available in kits of 1-12 and 13-27 duplex (79800 1 KT and 79800 2 KT).

Kit components:This kit is designed for input sheared genomic DNA amounts from 100 ng to 1 μg and consists of 3 tubes: Prep2Seq NGS Enzyme Mix, Prep2Seq NGS 2X Buffer, and Prep2Seq NGS High Concentration

Ligase. A streamlined workflow, straightforward pro-tocol, and high performing kit ensures quality library prep for your Illumina® sequencing runs.

Per 10 reactions: Prep2Seq NGS Enzyme Mix, 40 μl Prep2Seq NGS 2X Buffer, 250 μl Prep2Seq NGS High Concentration Ligase, 40 μl


WorkflowTime Comparison


min Ligation set up

NEB Next Ultra

USB Prep2Seq 5

min Ligation set up

30min Incubation

40min Incubation


Incubation55min

Clean up

5min

End repair set up

30min Incubation

35min

Clean up

5 30min Incubation

5

5 10min Incubation

90min Clean up

30min Clean up


min A-tail set up





min Ligation set up



78500 100 reactions

The Thermo Sequenase Cycle Sequencing Kit allows for equally efficient incorporation of both ddNTPs and dNTPs in cycle sequencing reactions resulting in very uniform band intensities which can be read accurately. The kit offers the choice of a three-dNTP extended primer method with internal α-labeled dNTPs or a 5' end-labeled primer method (see Table 1).

Obtain high quality sequenceLike Sequenase DNA Polymerase, Thermo Sequenase generates sequences with uniform band intensities and does not have the variability in band intensities of Taq DNA polymerase(1,2).

Flexible labelingThe kit can be used for either internal labeling with α-labeled dNTPs or with 5' end-labeled primers.

Use less templateAs little as 5 fmol of single-stranded or 20 fmol of double-stranded template is needed.

Read through secondary structureRegions with palindromes, hairpins or base repeats are more easily resolved with Thermo Sequenase and cycle sequencing methods.

Resolve gel compressions with 7-deaza-dGTP nucleotide mixesWhen DNA is synthesized with 7-deaza-dGTP instead of dGTP, weaker secondary structures are formed, thus reducing compression artifacts in sequencing gels.

Eliminate weak bands with pyrophosphataseOccasional weak bands occur with prolonged reac-tion or cycle times(3,4). This is caused by sequence-specific pyrophosphorolysis which is catalyzed by the polymerase. The addition of pyrophosphatase minimizes pyrophosphorolysis and can prevent incor-rect base-calling due to weak band intensities.

Kit components:Thermo Sequenase with thermostable

pyrophosphataseControl DNA pUC18Control Primer (-40 Forward, 23-mer)

Cycle Mix 7-deaza-dGTPCycle Mix dATPCycle Mix dTTPCycle Mix dCTP

ddGTP Termination Mix (7-deaza-dGTP)ddATP Termination Mix (7-deaza-dGTP)ddTTP Termination Mix (7-deaza-dGTP)ddCTP Termination Mix (7-deaza-dGTP)

Reaction Buffer (concentrate)Stop Solution


Important notes:The formulation of Thermo Sequenase DNA Polym-erase in this kit requires the use of a glycerol tolerant DNA sequencing gel to eliminate glycerol-induced distortion of bands at approximately 350 to 600 bases beyond the primer. Information on use is included in the kit protocol. If this region is beyond the reading range, Tris-Borate-EDTA Buffer (TBE) may be used.

References:1. Tabor, S. and Richardson, C. C. (1995) Proc.Natl.

Acad.Sci.USA 92, 6339-6343.2. Samols, S. B., McArdle, B. F., Ruan, C. C.,

Vanderhorn, P. B. and Fuller, C. W. (1995) Comments 22, No. 2, United States Biochemical Corp., Cleveland, OH.

3. Ruan, C. C., Samols, S. B. and Fuller, C. W. (1990) Comments 17, No.1, United States Biochemical Corp., Cleveland, OH.

4. Tabor, S. and Richardson, C. C. (1990) J.Biol.Chem. 265, 6447-6458.

Related products

ExoSAP-IT® PCR Product Cleanup78250 20 reactions78200 100 reactions78201 500 reactions

PrepEase® Quick MiniSpin Plasmid Kit78742 250 preps




DNA Sequencing | Thermo Sequenase Kits

Thermo Sequenase Cycle Sequencing Kit

Fig. 1. Two step cycle sequencing of plasmid DNA using the Thermo Sequenase Cycle Sequencing Kit and α-33P-dATP.

G A T C

Table 1. Cycle sequencing requirements

MethodLabel useda

Sensitivityb μg (fmol) M13

Special requirements

Typical use

Total timec

3-dNTPInternal labelcycle protocol

5µCiα-dATP

≥ 0.01 μg (5 fmol)

Must design primer for cycled labeling.

Direct sequencing of smallest quantities of less-pure DNA.

3–4 hours

Radiolabeled primer cycle protocol

γ-dATP ≥ 0.01 µg (5 fmol) (33P)

T4 Polynucleotide Kinase, γ-ATP for labeling primer.

Repeated sequencing of similar templates.

2–3 hours

a In most cases, internal (α-dATP) labeling can be performed using 35S, 33P, or 32P-labeled dATP or dCTP.b Nominal minimum quantities of M13 DNA giving clear, reliable sequence with overnight exposure on film using 33P labeled nucleotide.c Including cycling time. Hands-on time is approximately 30 - 40 minutes for all methods.


DNA Sequencing | Thermo Sequenase Kits

79260 50 reactions

The Thermo Sequenase Dye Primer Manual Cycle Sequencing Kit is designed to be used with fluorescent dye-labeled primers and high-resolution fluorescence scanners. Sequencing reactions involve a termination step followed by standard slab gel electrophoresis and scanning of the gel to detect fluorescently tagged sequencing products.

Direct sequencingSequence from bacterial colonies, M13 or lambda plaques. Single yeast patches on plates or small liquid culture may be sequenced by using this kit combined with the PrepEase Yeast Plasmid Isolation Kit (PN 79220).

Single-step protocolSequencing reactions involve use of fluorescent-labeled primers and a termination step. Note: The choice of fluorescent dyes depends on the scanner. For instruments equipped with a 532 nm laser, TAMRA (Rhodamine dye), HEX (fluorescein dye), ROX, and FAM labeled primers may be used.

Flexible detectionThe gel can be scanned without opening the glass plates, thus, allowing gel electrophoresis to be stopped at any point for analysis and then resumed if longer read lengths are needed.

Thermo sequenase DNA polymerase/tap enzyme mixThe enzyme is a mixture of Thermo Sequenase and thermostable inorganic pyrophosphatase (TAP) cloned from the thermophile Thermoplasmaacidophilum. TAP prevents pyrophosphorolysis from occurring. Pyrophosphorolysis can result in faint bands, which can affect the accuracy of base calling.

Resolve gel compressions with 7-deaza-dGTP nucleotide mixesWhen DNA is synthesized with 7-deaza-dGTP instead of dGTP, weaker secondary structures are formed, thus reducing compression artifacts in sequencing gels.

Specialized formamide loading dyeA special formulation of the two tracking dyes does not obscure TAMRA or HEX primers. The faster migrating dye fluoresces at a wavelength greater than 649 nm. The slower migrating dye fluoresces at a wavelength greater than 600 nm. Thus, sequences will not be obscured if TAMRA or HEX labeled prim-ers are used.

Direct loading of samples on the gel Alcohol precipitation of the sequencing reaction products is not required. Simply load the reaction onto a standard sequencing gel.

Important notes:The formulation of Thermo Sequenase DNA Polymerase in this kit necessitates the use of a glycerol tolerant DNA sequencing gel to eliminate glycerol-induced distortion of bands at approximate-ly 350 to 600 bases beyond the primer. Information on use is included in the kit’s protocol. If this region is beyond the reading range, Tris-Borate-EDTA Buffer (TBE) may still be used.

Kit components:Thermo Sequenase with thermostable inorganic

Pyrophosphatase Reaction Buffer (concentrate) Control DNA pUC19Control Primer, TAMRA -40 Universal Forward PrimerddG Termination Mix (7-deaza-dGTP)ddA Termination Mix (7-deaza-dGTP)ddT Termination Mix (7-deaza-dGTP)ddC Termination Mix (7-deaza-dGTP)Formamide Loading Dye


Related products

ExoSAP-IT PCR Product Cleanup78250 20 reactions78200 100 reactions78201 500 reactions


PrepEase BAC Purification Kit78722 10 preps

PrepEase Plasmid 8-well Strip Kit78745 12 x 8 strips




PrepEase Yeast Plasmid Isolation Kit79220 50 preps

Thermo Sequenase Dye Primer Manual Cycle Sequencing Kit

Fig. 1. Direct sequencing from single bacterial colony using the Thermo Sequenase Dye Primer Manual Cycle Sequencing Kit.

250

200

150

100

G A T C


DNA Sequencing | Sequenase Polymerase


Sequenase Version 2.0 Reaction Buffer, 5X(Included with each pack size of enzyme)70702 1 ml

Source:E.colistrain containing clones that overproduce T7 Gene 5 Protein and E.coliThioredoxin.

Description:Sequenase Version 2.0 DNA Polymerase is a genetically engineered form of T7 DNA polymerase(1). Unlike the wild-type enzyme, it has virtually no 3'→5' exonuclease activity. Sequenase Version 2.0 is highly processive, incorporates nucleotide analogs (dlTP, thio-dNTPs, dideoxy-NTPs, etc.), is not impeded by secondary structures and can carry out strand displacement synthesis. It is an excellent enzyme for dideoxy-sequencing and is useful in other applica-tions, especially where the absence of associated exonuclease activity is desirable.

Applications:�� DNA Sequencing�� Second strand synthesis for microarray applications(2)

�� Random primed labeling with nucleotide analogs(2)

�� Labeling the 3' termini of DNA fragments with 5' protruding ends�� Amplification of DNA�� Strand displacement applications

Note: Sequenase Version 2.0 DNA Polymerase may not be useful for making blunt-end fragments because it can leave a one-base over-hang at the 3' end.

Properties:Molecular Weight: Consists of two subunits,

modified T7 gene 5 protein (76 kDa) and E.coli thioredoxin (12 kDa).

Optimum pH: 7.5Optimum Temperature: 37°CRequirements for Divalent Cation: Mg2+, Mn2+

Purity:Greater than 95% pure as determined by SDS-PAGE. Tested for contaminating double- and single-strand-ed endonucleases and exonucleases.

Storage buffer:20 mM potassium phosphate (pH 7.4), 1 mM DTT, 0.1 mM EDTA, 50% glycerol.

Assay conditions:The reaction mixture (100 µl) contains 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 5 mM DTT, 0.3 mM dNTPs and 5 µg M13mp18 pre-annealed to 5 pmol M13 universal primer; incubation is at 37°C for 1 minute.

Unit definition:One unit of enzyme catalyzes the incorporation of 1 nmol of nucleotide into acid insoluble form in 30 seconds at 37°C.


Functional test:DNA sequencing with the Sequenase Version 2.0 DNA Sequencing Kit (PN 70770).

Functionally Tested 5X Sequenase Version 2.0 Reaction Buffer (1 ml included, PN 70702):200 mM Tris-HCl (pH 7.5), 100 mM MgCl2, 250 mM NaCl

Sequenase Version 2.0 Dilution Buffer (1 ml included):10 mM Tris-HCl (pH 7.5), 5 mM DTT, 0.1 mM EDTA


References:1. Tabor, S. and Richardson, C. C. (1989) J.Biol.

Chem. 264, 6447-6458.2. Wang, D., Coscoy, L., Zylberberg, M., Avila, P. C.,

Boushey, H. A., Ganem, D. and DeRisi, J. L. (2002) Proc.Natl.Acad.Sci.USA, 99, 15687-15692.

3. Paris, M. (1992) Comments18, (No. 3), United States Biochemical Corporation, Cleveland, Ohio.

4. Tabor, S. and Richardson, C. C. (1987) Proc.Natl.AcadSci.USA84, 4767-4771.

5. Tabor, S. and Richardson, C. C. (1989) Proc.Natl.Acad.Sci.USA86, 4076-4080.

Related products

Control DNA, M13mp18, 0.2 µg/µl70704 50 µl 50 µg

DTT Solution, 0.1 M70726 150 µl

Labeling Mix for dGTP, 5X70710 100 μl

Mn Buffer72600 100 μl

Stop Solution70724 1,200 µl

ddA Termination Mix for dGTP70714 250 μl

ddA Termination Mix for dITP70728 125 µl

ddC Termination Mix for dGTP70716 250 μl

ddG Termination Mix for dGTP70718 250 μl

ddT Termination Mix for dGTP70720 250 μl

Sequenase Version 2.0 DNA Polymerase (E.C.2.7.7.7)


DNA Sequencing | Sequenase Kits

70770 100 reactions

The Sequenase Version 2.0 DNA Sequencing Kit features Sequenase Version 2.0 DNA Polymerase, the standard for high quality DNA sequencing(1). Sequenase Version 2.0 DNA polymerase is a genetically engineered form of T7 DNA polymerase which retains extraordinary polymerase activity with virtually no 3'→5' exonuclease activity. It is highly processive, able to effectively incorporate nucleotide analogs for sequencing (dideoxy NTPs, α-thio dATP, dITP, 7-deaza-dGTP, etc.) and is not easily impeded by template secondary structure. The kit includes all the reagents necessary to achieve high quality results (Fig. 1). Use of the specially formulated buf-fers and mixes included in the kit will maximize yield of sequence information.

Flexible labelingThe kit can be used for either internal labeling with α-labeled dNTPs or with 5'end-labeled primers.

Resolve gel compressions with dITP nucleotide mixesThe substitution of dITP (deoxyinosine triphosphate) for dGTP in the reaction mix eliminates the second-ary structures that produce gel compressions. dITP forms fewer H-bonds with dCTP than does dGTP, so product is more readily denatured during gel electrophoresis. Hence, sequence data is free from gel-based compression artifacts and results are more accurate.

Emphasize sequence close to primers with Mn bufferManganese (Mn) is added to emphasize sequence close to the primer, which may be weak if insuf-ficient DNA template is used. Mn++ increases the incorporation rate of dideoxynucleotides relative to deoxynucleotides. Thus, termination occurs earlier and more sequence is visible close to the primer(2,3).

Increase read length with sequence extending mixesThese mixes enable chain terminations to be extended to more than 3,000 bases from the primer. The use of these mixes provides a simple method to further extend the range of sequence, if needed. Keep in mind that this degree of extension can reach well beyond the limits of any electrophoresis gel resolution, yet use of the mixes when combined with short and long gel runs can increase overall sequence yield.

Eliminate weak bands with pyrophosphataseOccasionally, weak bands may occur with prolonged reaction times (greater than 5 minutes)(4,5), or when dITP is used in the sequencing reaction(6). The addition of pyrophosphatase can prevent weak band intensities brought on by sequence-specific pyrophosphorolysis catalyzed by the polymerase.

Convenient enzyme storage with glycerol enzyme dilution bufferSequenase DNA polymerase can be pre-diluted to a working concentration for storage of the enzyme in this form. This eliminates the necessity of diluting the polymerase prior to each sequencing reaction. Also, the addition of glycerol enhances the stability of the enzyme in sequencing reactions. Note: Pre-dilution of the polymerase with this buffer results in higher glycerol concentration in the sequencing reaction. A Glycerol Tolerant Gel (GTG) Buffer (PN 75827) must be used in the sequencing gel and buffer chambers to eliminate glycerol-induced distortion of bands at approximately 350 to 600 bases beyond the primer(7). If this region is beyond the region of inter-est, Tris-Borate-EDTA (TBE) Buffer may be used (PN 70454 or 75891).

Kit components:Sequenase Version 2.0 DNA PolymeraseInorganic Pyrophosphatase

Enzyme Dilution BufferGlycerol Enzyme Dilution BufferSequenase Reaction Buffer (5X)Dithiothreitol SolutionMn Buffer

Control DNA M13 mp18Primer (-40 Universal)

Labeling Mix (dGTP, 5X)ddGTP Termination Mix (for dGTP)ddATP Termination Mix (for dGTP)ddTTP Termination Mix (for dGTP)ddCTP Termination Mix (for dGTP)Sequencing Extending Mix (for dGTP)

Labeling Mix (dITP, 5X)ddGTP Termination Mix (for dITP)ddATP Termination Mix (for dITP)ddTTP Termination Mix (for dITP)ddCTP Termination Mix (for dITP)Sequence Extending Mix (for dITP)

Stop Solution


References:1. Tabor, S. and Richardson C. C. (1989) J.Biol.

Chem. 264, 6447-6458.2. Fuller, C. W. (1989) Comments 16, No. 3, United

States Biochemical Corp., Cleveland, OH.3. Tabor, S. and Richardson, C. C. (1989) Proc.Nat.

Acad.Sci.USA 86, 4076-4080.4. Ruan, C. C., Samols, S. B. and Fuller, C. W. (1990)

Comments 17, No. 1, United States Biochemical Corp., Cleveland, OH.

5. Tabor, S. and Richardson C. C. (1990) J.Biol.Chem. 265, 8322-8328.

6. Tabor, S. and Richardson, C. C. (1989) Proc.Nat.Acad.Sci.USA 84, 4767-4771.

7. Pisa-Williamson, D. And Fuller, C. W. (1992) Comments 19, No. 2, United States Biochemical Corp., Cleveland, OH.

Sequenase Version 2.0 DNA Sequencing Kit

Fig. 1. Standard sequencing results using 35S-labeled dATP and the Sequenase Version 2.0 DNA Sequencing Kit.


70170 100 reactions

The Sequenase PCR Product Sequencing Kit utilizes a novel enzymatic cleanup method to pre-treat PCR products prior to sequencing without any subsequent purification or separation steps. This kit contains all of the standard Sequenase Version 2.0 Kit components with the addition of Shrimp Alkaline Phosphatase (SAP) and Exonuclease I to effectively remove excess dNTPs and primers present in the final PCR product reaction mixture. Both enzymes are added simultaneously to a specific aliquot of the PCR product mixture, incubated at 37°C and then inactivated at 80°C. The result is a cleaner PCR product, free from excessive nucleotides and primers, ready to be sequenced with standard Sequenase reagents.

Flexible labelingThe kit can be used for either internal labeling with α-labeled dNTPs or with 5'end-labeled primers.

Resolve gel compressions with 7-deaza-dGTP nucleotide mixesWhen DNA is synthesized with 7-deaza-dGTP instead of dGTP, weaker secondary structures are formed, thus reducing compression artifacts in se-quencing gels. Generally, 7-deaza-dGTP is preferred over dITP since band signal tends to be stronger and more uniform. However, dITP is still recommended for resolving the strongest gel compressions.

Eliminate weak bands with pyrophosphataseOccasionally, weak bands may occur with prolonged reaction times (greater than 5 minutes)(3,4), or when dITP is used in the sequencing reaction(5). The addition of pyrophos phatase can prevent weak band intensities brought on by sequence-specific pyrophosphorolysis catalyzed by the polymerase. The formulation of Sequenase enzyme included in this kit is pre-mixed with pyrophosphatase for added convenience.

Convenient use of sequenase enzymeSequenase Version 2.0 DNA Polymerase within this kit is pre-diluted to a working concentration. This eliminates the necessity of diluting the polymerase prior to each sequencing reaction. Also, the enzyme is stored in 50% glycerol which enhances the stabil-ity of the enzyme in the sequencing reaction.

Kit components:Sequenase Version 2.0 DNA Polymerase with

Inorganic PyrophosphataseSAP, 200 unitsExonuclease I, 1,000 units

Sequenase Reaction Buffer (5X)Dithiothreitol SolutionMn Buffer

Control DNA (M13 clone)Primer (-40 Forward 23-mer)Primer (-50 Reverse 21-mer)dGTP, 3 µM dATP, 3 µMdTTP, 3 µM dCTP, 3 µM7-deaza-dGTP, 3 µM

Labeling Mix (dGTP, 5X)ddGTP Termination Mix (for dGTP)ddATP Termination Mix (for dGTP)ddTTP Termination Mix (for dGTP)ddCTP Termination Mix (for dGTP)

Labeling Mix (7-deaza dGTP, 5X)ddGTP Termination Mix (for 7-deaza-dGTP)ddATP Termination Mix (for 7-deaza-dGTP)ddTTP Termination Mix (for 7-deaza-dGTP)ddCTP Termination Mix (for 7-deaza dGTP)

Stop Solution


References:1. Fuller, C. W. (1989) Comments 16, No. 3, United

States Biochemical Corp., Cleveland, OH.2. Tabor S. and Richardson, C. C. (1989) Proc.Nat.



4. Tabor S. and Richardson, C. C. (1990) J.Biol.Chem. 265, 6447-6458.

5. Tabor S. and Richardson, C. C. (1987) Proc.Natl.Acad.Sci.USA 84, 4767-4771.

Sequencing kits

PCR Product Pre-Sequencing Kit70995 100 reactions70996 500 reactions70997 2,000 reactions

Sequenase Version 2.0 DNA Sequencing Kit70770 100 reactions

Sequenase Quick Denature PlasmidSequencing Kit70140 100 reactions

Thermo Sequenase Cycle Sequencing Kit 78500 100 reactions

Sequencing enzymes

Sequenase Version 2.0 DNA Polymerase 70775Y 200 units70775Z 1,000 units

PCR cleanup


HT ExoSAP-IT High-Throughput PCR Product Cleanup78395 480 reactions x 8-tube strip

5,760 reactions x 1 plate (12 x 8-tube strips)

23,040 reactions - 4 plates

1,000 reactions

5,000 reactions

Ultrapure reagents for DNA sequencing

Ammonium Persulfate76322 10 gm 100 gm 1 kg

Glycerol Tolerant Gel Buffer, 20X solution75827 1 L

RapidGel-40% Liquid Acrylamide75848 500 ml

RapidGel-XL-6%, TBE75861 500 ml

TEMED76320 100 gm 500 gm

TBE Buffer, 10X Powder70454 6 bottles

Sequenase PCR Product Sequencing Kit



70140 100 reactions

The Sequenase Quick Denature Plasmid Sequenc-ing Kit offers two simple and efficient denaturation methods that do not require precipitation of plasmid DNA. Both methods, Glycol/Glycerol and Alkali/Acid, allow for rapid generation of denatured template DNA suitable for use with standard Sequenase Version 2.0 DNA Polymerase. In addition, this kit contains the 7-deaza-dGTP nucleotide mixes which provide stronger band intensities for resolving sequencing gel compressions.

Rapid and simple denaturation protocolNo ethanol precipitation is needed. Simply denature plasmid template for up to 10 minutes, anneal primer for another 10 minutes and sequence.

Use less templateAs little as 0.5 µg of plasmid DNA is all that is needed to generate good sequence results.

Resolve gel compressions with 7-deaza-dGTP nucleotide mixes

Emphasize sequence close to primers with Mn bufferMn++ increases the incorporation rate of dide-oxynucleotides relative to deoxynucleotides. Thus, termination occurs earlier and more sequence is visible close to the primer(2,3).

Increase read length with sequence extending mixesThese mixes enable chain terminations to be extended to more than 3,000 bases from the primer. The use of these mixes provides a simple method to further extend the range of sequence, if needed. Keep in mind that this degree of extension can reach well beyond the limits of any electrophoresis gel resolution, yet use of the mixes when combined with short and long gel runs can increase overall sequence yield.

Eliminate weak bands with pyrophosphataseOccasionally, weak bands may occur with prolonged reaction times (greater than 5 minutes)(3,4), or when dITP is used in the sequencing reaction(5). The addition of pyrophos phatase can prevent weak band intensities brought on by sequence-specific pyrophosphorolysis catalyzed by the polymerase. The formulation of Sequenase enzyme included in this kit is pre-mixed with pyrophosphatase for added convenience.

Convenient use of sequenase enzymeSequenase Version 2.0 DNA Polymerase within this kit is pre-diluted to a working concentration. This eliminates the necessity of diluting the polymerase prior to each sequencing reaction. Also, the enzyme is stored in 50% glycerol which enhances the stabil-ity of the enzyme in the sequencing reaction.

Kit components:Sequenase Version 2.0 DNA Polymerase with

inorganic pyrophosphatasePlasmid Reaction Buffer (concentrate)Plasmid Denaturing ReagentNaOH, 1.0 M SolutionHCl, 1.0 M SolutionDithiothreitol SolutionMn Buffer

Control DNA pUC19Primer (-40 Forward, 23-mer)Primer (-50 Reverse, 21-mer)

Labeling Mix (7-deaza-dGTP, 5X)ddGTP Termination Mix (7-deaza-dGTP)ddATP Termination Mix (7-deaza-dGTP)ddTTP Termination Mix (7-deaza-dGTP)ddCTP Termination Mix (7-deaza-dGTP)

Stop Solution


References:1. Fuller, C. W. (1989) Comments 16, No. 3, United

States Biochemical Corp., Cleveland, OH.2. Tabor S. and Richardson, C. C. (1989) Proc.Nat.



4. Tabor S. and Richardson, C. C. (1990) J.Biol.Chem. 265, 6447-6458.

5. Tabor S. and Richardson, C. C. (1987) Proc.Natl.Acad.Sci.USA 84, 4767-4771.

6. Pisa-Williamson, D. and Fuller, C. W. (1992) Comments 19, No. 2, United States Biochemical Corp., Cleveland, OH.

Sequencing kits


Sequenase PCR Product Sequencing Kit70170 100 reactions

Thermo Sequenase Cycle Sequencing Kit78500 100 reactions

Sequencing enzymes

Sequenase Version 2.0 DNA Polymerase 70775Y 200 units70775Z 1,000 units

Sequencing reagents

Glycerol Tolerant Gel Buffer, 20X solution75827 1 L

RapidGel-40%75848 500 ml

RapidGel-XL-6%75861 500 ml

Taurine, Ultrapure75824 1 kg

Sequenase Quick Denature Plasmid DNA Sequencing Kit



DNA Sequencing | Sequenase Primer/Sequencing Tips

Primer for DNA Sequencing

T3 Promoter Primer

Prod. No. Product Description Primer Sequence [conc] Size

71200 T3 Promoter 5' ATTAACCCTCACTAAAG 3' 5 pmol/µl 250 pmol

Determining Primer Concentration

Approximate extinction coefficients and molecular weights for oligonucleotides

DNA Sequencing Gel Formulations

Standard sequencing gels Acrylamide/ Urea 20X Glycerol 10X TBE

Gel Conc. Bis-Acrylamide (7.0 – 8.3 M) Tolerant Gel Buffer Buffer H2O

6% 5.7 gm/0.3 gm 42 – 50 gm 5 ml** - ~ 45 ml

8% 7.6 gm/0.4 gm 42 – 50 gm 5 ml** - ~ 45 ml

6% 5.7 gm/0.3 gm 42 – 50 gm - 10 ml ~ 40 ml

8% 7.6 gm/0.4 gm 42 – 50 gm - 10 ml ~ 40 ml

Dissolve, adjust volume to 100 ml with H2O, filter and de-gas. When ready to pour, add 1 ml of 10% ammonium persulfate and 25 µl TEMED (N, N, N', N' tetrameth-ylethylene-diamine).

** Use 4 ml for faster gel migration.

Formamide sequencing gels Acrylamide/ Urea 20X Glycerol 10X TBE Gel Conc. Bis-Acrylamide (7 M) Tolerant Gel Buffer Buffer Formamide H2O

6% 5.7 gm/0.3 gm 42 gm 5 ml - 30 – 40 ml ~ 15 ml

8% 7.6 gm/0.4 gm 42 gm 5 ml - 30 – 40 ml ~ 15 ml

6% 5.7 gm/0.3 gm 42 gm - 10 ml 30 – 40 ml ~ 15 ml

8% 7.6 gm/0.4 gm 42 gm - 10 ml 30 – 40 ml ~ 15 ml

Warming to 35 - 40°C may be required to dissolve completely. Adjust volume to 100 ml with H2O, filter and de-gas. When ready to pour, add 1 ml of 10% am-monium persulfate and 100 - 150 µl TEMED. These formamide-containing gels may be used for very strong compressions not resolved by 7-deaza-dGTP. They will require higher running voltage and run slower than urea-only gels. Prior to drying, these gels should be soaked in 5% acetic acid, 20% methanol to prevent swelling.

N = # of bases

ε260 ≅ 10,000 x N(M-1 cm-1)

Molecular Weight ≅ 330 x N

(OD260 ÷ ε260) x 106 = Concentration (µM)

Concentration (µM) x Molecular Weight = Concentration (ng/ml)

Example for a 17 mer: ε260 ≅ 170,000 (M-1 cm-1) Molecular Weight ≅ 5,600

A solution with OD260 = 1.0 will be: (1.0 ÷ 170,000) x 106 = 5.9 µM 5.9 x 5,600 = 33,000 ng/ml


DNA Sequencing | Sequencing Tips

DNA Sequencing Gel Formulations continued...

General guidelines for running polyacrylamide sequencing gels(Based on 100 ml gel solutions for regular, wedge and formamide gels) 40% Regular Wedge Formamide

Acrylamide:Bis-Acrylamide 19:1 19:1 19:1

10% Ammonium Persulfate 1 ml 1 ml 1 ml

TEMED 30 µl 30 µl 150 µl

Gel Running 55 Watts 55 Watts 55 Watts

Conditions 12 – 1600V 12 – 1600V 19 – 2500V

(Constant Power) ~40 mA ~40 mA ~25 mA

Secondary structure in DNA templates is a common cause of BAFLs (Bands Across Four Lanes observed in sequencing gels), also referred to as Stops. When Sequenase Version 2.0 Polymerase encounters a stable secondary structure in the template, it may dissociate from the template without incorporating a dideoxynucleotide. The fragments generated will co-migrate and cause artifact bands.

BAFLs can be eliminated by performing a chase step with TdT to remove artificially terminated fragments. Terminal Deoxynucleotidyl Transferase is a polymerase which non-specifically adds nucleotides to a free 3' hydroxyl. Its action in this case is to extend the fragments which are not properly terminated with a dideoxynucleotide. TdT will add hundreds of nucleotides in a relatively short period of time allowing non-specifically terminated fragments to be extended such that they run at the top of the gel without interfering with readable sequence.

This method allows for the complete resolution of BAFLs and difficult gel compressions.

Method1. A 1 mM solution of all four dNTPs is made in Sequenase Enzyme Dilution

Buffer. 1 µl each dNTP (100 mM stock) 96 µl Enzyme Dilution Buffer2. Terminal Deoxynucleotidyl Transferase (TdT) [PN 72033] is diluted in the

1 mM dNTP solution (above) to 2 units/µl. Each termination reaction will require 1 µl of this enzyme/dNTP dilution. Mix well on ice. It is best to prepare this enzyme/dNTP solution immediately prior to use.

3. Sequence according to the Sequenase Version 2.0 Sequencing Protocol using the appropriate labelling and termination mixes. DO NOT add Stop Solution.

4. After the 2-5 minute termination reaction is complete add 1 µl of the TdT/dNTP solution to each termination reaction tube.

5. Mix and incubate 15-30 minutes at 37°C.6. Add 4 µl of Stop Solution to each reaction tube.7. Heat denature and load on a sequencing gel as usual.

References:1. Tabor, S. and Richardson, C. C. (1987) Proc.Natl.Acad.Sci USA 84, 4767-

4771.2. Fawcett, T. W. and Bartlett, S. G. (1991) Comments, (United States

Biochemical Corporation), 17, (4) pg. 19.3. Fawcett, T. W. and Bartlett, S. G. (1990) BioTechniques 9, 46-49.

Related products

dNTP 100 mM Solutions, Set of Four4 x 25 µmol (0.25 ml) each dNTP per pack77100 1 pack


Terminal Deoxynucleotidyl Transferase, Recombinant (rTdT)72033 500 units 2,500 units

Thermo Sequenase Cycle Sequencing Kit78500 100 reactions

Use of Terminal Deoxynucleotidyl Transferase (TdT) to Resolve Bands Across Four Lanes (BAFLs)

Fig. 1. Use of TdT to eliminate BAFLs. Lane A: Plasmid sequence with 2 strong BAFLs indicated by arrows. Lane B: Sequence results with same plasmid that included a TdT/dNTP chase reaction.

A B



Compression artifacts may be produced during electrophoresis due to the formation of stable secondary structures in the sequencing product DNA fragments. The folded structures run faster through the gel matrix than equivalent unfolded DNA fragments; consequently they will migrate to the same position as smaller sized fragments resulting in bands across 2 or more lanes. Typical indications of a compression are odd spacing, bands very close together followed by bands spaced farther apart than normal, or bands in two to four lanes at the same point. Sometimes bases can be lost entirely. The most difficult instances are those in which the compressed area looks perfectly normal. These are detected when the opposite strand is sequenced or nucleotide analogs are used to check for compression artifacts. Nucleotide analogues such as 7-deaza-dGTP or dITP can be used to alleviate compressions but in some instances are not able to fully resolve a difficult compression. In these cases, the use of a formamide gel alone or in conjunction with the nucleotide analogue, will completely relieve the compressed areas giving unambiguous sequence.

MethodPreparation and running a formamide gel is not very different from a standard sequencing gel. A 6% or 8% acrylamide gel may be prepared. The running time is increased to approximately one and one-half times that of a standard gel.

1. Weigh the following dry ingredients into a beaker:6% gel 5.7 gm Acrylamide

0.3 gm N-N' Methylene-bis-acrylamide 42 gm Urea

8% gel 7.6 gm Acrylamide 0.4 gm N-N' Methylene-bis-acrylamide 42 gm Urea

2. Add: 40 ml Formamide 10 ml 10X TBE Buffer OR 5 ml 20X GTG Buffer (for a Glycerol Tolerant Gel)

and a magnetic stir bar

3. Warm the gel mix to 50°C and stir until the components are completely dissolved.

4. Continue stirring until the temperature cools to ~30°C. Adjust the volume to 100 ml with distilled water and vacuum filter with a nitrocellulose filter unit. The gel should be poured immediately. To prevent the urea from precipitating, do not allow the gel to cool below 25°C. If the mix should happen to precipitate the process must be repeated from the beginning; however running a gel that has been redissolved will give very poor results.

5. Add 1 ml of 10% ammonium persulfate and 25 µl of TEMED. (NOTE: Use this amount of TEMED when pouring the gel the night before it is to be run. If the gel is to be used the same day, adding four to five times the amount of TEMED will polymerize the gel in an hour. It is usually more convenient to pour the gel at the end of one day and run it first thing the next day to accommodate the longer running time of formamide gels). Mix well and pour immediately into clean gel plates. The solution is quite viscous and somewhat more difficult to pour than a standard gel. It may be necessary to tip the plates completely upright to get the gel mix to spread properly between them.

6. When preparing the gel for electrophoresis, be sure to clean out the wells thoroughly prior to loading the samples. Run the gel at 60W constant power. Expect the voltage to climb to 2,000V and the plates to reach a higher temperature than usual as the gel progresses. Normally the dye fronts eventually run as thin bands.

7. Stop the gel at the desired point and soak it in a 10% acetic acid, 20% methanol solution for 45 minutes or until shrinkage is apparent. Formamide gels tend to swell when placed into the soaking solution, however, with time they will shrink to a manageable size. Dry and expose the gel as you would any other. In some cases the formamide gel may require slightly longer drying time than a standard gel.

Relief from Compression Artifacts in DNA Sequencing Using Formamide Gels

Fig. 1. Sequence determination using Sequenase Version 2.0 DNA Polymerase of the single-stranded vector M13mp18. A primer at position 6511 is used to visualize a very signifi-cant gel compression. A. Sequence determined using dGTP and 7-deaza-dGTP nucleotide mixes and run on a 6% TBE gel. Note the significant compression in both reactions. The sequence is unreadable at this point. B. The same reactions run on an 8% TBE/40% Formamide gel. Note the G doublet is not completely resolved in the dGTP reaction even on this Formamide gel while the 7-deaza-dGTP reaction is completely resolved.

A B

7-deaza dGTP dGTP dGTP

7-deaza dGTP

Related products


Ammonium Persulfate (APS)76322 10 gm 100 gm

bis-Acrylamide75821 25 gm 50 gm 100 gm

Formamide75828 500 gm

1 kg

TEMED76320 100 ml

Urea75826 1 kg

5 kg



Prevent Weak Sequencing Band Intensities by Using Pyrophosphatase

The determination of DNA sequence using a DNA polymerase (Klenow) and nucleotide-specific chain-termination was first described by F. Sanger etal.(1). In this procedure, DNA synthesis is initiated at a unique priming site, and terminated by the incorporation of 2',3'-dideoxynucleoside monophosphates (ddNMPs). Tabor and Richardson(2) noted that under certain conditions, the intensities of some of the bands on resulting sequencing gels can be weak. This phenomenon is particularly apparent when the reactions are run for 30 minutes or longer and when dITP is used in place of dGTP. These weak bands are the result of pyrophosphorolysis(3,4). This is the reversal of the polymerization reaction catalyzed by the DNA polymerase (see below). Pyrophosphorolysis can be prevented using inorganic pyrophosphatase to hydrolyze the pyrophosphate(3,4).

Reactions of DNA PolymerasesAll known DNA polymerases catalyze the template-dependent incorporation of a deoxynucleotide onto the 3' hydroxyl terminus of a primer, with the re-lease of inorganic pyrophosphate (PPi). Catalysis requires the presence of a divalent cation, either Mg2+ or Mn2+. Under the conditions normally used for DNA synthesis (i.e., high dNTP concentrations and low pyrophosphate con-centration), the forward reaction (polymerization) is greatly favored over the reverse reaction (pyrophosphorolysis). Many DNA polymerases also catalyze the hydrolysis of nucleotides from the 3' terminus of DNA through a sepa-rate exonuclease activity. This reaction is unlike pyrophosphorolysis because the substrate is H2O and the product is a nucleoside monophosphate.

The polymerization, pyrophosphorolysis and exonuclease reactions are sum-marized below:

M2+

Polymerization: DNA(n) + dNTP → DNA(n+1) + PPi

M2+

Pyrophosphorolysis: DNA(n) + dNTP ← DNA(n+1) + PPi

M2+

Exonuclease Reaction: DNA(n) + H2O → DNA(n-1) + dNMP (M2+ = Mg2+ or Mn2+)

Although pyrophosphorolysis is much slower than polymerization, it can affect DNA sequencing reactions(3,4). Some bands become less intense when the termination reactions are allowed to incubate for extended periods of time. When the reactions are incubated for 2 minutes, the bands have uni-form intensities; however, after a 30 minutes incubation, approximately one in ten bands is reduced in intensity. Note that the rates at which individual bands decrease in intensity varies widely, depending on the DNA sequence.

Sequence-specific decreases in band intensity occur even when sequenc-ing with DNA polymerases that have no exonuclease activity such as AMV Reverse Transcriptase and Sequenase Version 2.0 DNA Polymerase(5). Band weakening is stimulated by the addition of pyrophosphate to the sequenc-ing reactions and it is prevented by the addition of inorganic pyrophos-phatase or high concentrations of dNTPs(3,4). Thus, the decrease in intensity of some bands with longer incubation times is due to pyrophosphorolysis.

Action of Inorganic PyrophosphataseInorganic pyrophosphatase catalyzes the hydrolysis of pyrophosphate to two molecules of orthophosphate.

Mg2+

P2O74- + H2O → 2 HPO4

2-

This enzyme plays an important role in DNA synthesis invivo, rendering DNA polymerization essentially irreversible by removing pyrophosphate. The addition of inorganic pyrophosphatase to DNA sequencing reactions com-pletely stabilizes all band intensities, even when incubating the reactions for 60 minutes and when dITP is used in place of dGTP.

For convenience, the USB Sequenase Version 2.0 DNA Sequencing Kit (PN 70770) includes nuclease-free yeast inorganic pyrophosphatase. Pyrophosphatase can be used for sequencing with dGTP or its analogs (dITP, 7-deaza-dGTP, etc.) and is effective in the presence of Mg2+ or Mn2+. The use of both Sequenase polymerase and pyrophosphatase assures the stability of all DNA generated in the DNA sequencing reaction, providing additional assurance that the sequence determined is accurate.

References:1. Sanger, F., Nicklen, S. and Coulson, A. R. (1977) Proc.Natl.Acad.Sci.USA

74, 5463-5467.2. Tabor, S. and Richardson, C. C. (1987) Proc.Nat.Acad.Sci.Usa 84, 4767-

4771.3. Tabor, S. and Richardson, C. C. (1990) J.Biol.Chem. 265, 8322-8328.4. Ruan, C. C. Samols, S. B. and Fuller, C. W. (1990) Comments17, No. 1,

1-27.5. Tabor, S. and Richardson, C. C. (1989) J.Biol. Chem. 264, 6447-6458.

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BiochemicalsAcrylamides

ACS Biochemicals

Agaroses

Albumins

Amino Acids

Antibiotics

Bacterial Culture Media

Buffers

Enzymes

Proteins

Substrates

Sugars

Ultrapures

Vitamins

Biochemicals


Formulas and formula weights are provided for compounds having a specific known structure. The values used to calculate formula weights are taken from the 2001 Table of Atomic Weights, (2003) Pure Appl. Chem., 75, 1107-1122.

To simplify grade selection for customers the following grade definitions are provided. We have selected grade names which are self explanatory, as well as those which reflect various industry standards. If any changes are noticed in a familiar grade designation for a given chemical, it is a result of a change to the grade definition rather than a change in the quality of the chemical.

USB Ultrapures are specially selected biochemicals and reagents of high quality, suitable for use in numerous biotechnology applications. These re-agents meet stringent requirements, such as adherence to compendial refer-ences where applicable, have ultra-low levels of trace metal contamination, and low levels of DNase, RNase, protease, and endotoxins where applicable.

Reference constants for nucleotides and nucleosidesValues for reference constants are taken from the literature. They are useful in determining the concentration of stock solutions prepared in the laboratory.

DBR: “Data for Biochemical Research” Dawson, R. M. C., Elliot, D. C., Elliot, W. H. , Jones, K. M., (1986) Oxford Science Publications.

NRC: “Specifications and Criteria for Biochemical Compounds,” (1972) National Academy of Sciences, Washington, D.C. 3rd edition.

Biochemical grade definitionsACS Reagent Grade: Meets the limits of purity for inorganic chemicals as established by the American Chemical Society.

CP Grade (Chemically Pure): Meets the requirements of the Food Chemicals Codex, but not intended for food use.

Genetic Performance Certified (GPC): Functionally tested in specific molecular biology applications (e.g. nucleic acid recovery, in-gel restriction enzyme digestion or ligation) to ensure optimal agarose performance.

Hi-Res™: Agaroses especially selected for high resolution of samples.

USBioAnalyzed: Meets USP or NF chemical testing requirements but is not sold as USP or NF Grade; USP and NF Grades (United States Pharmacopeia and National Formulary) must meet the requirements of those compendia and will fall under this category.

USP Grade: Meets the requirements of the U.S. Pharmacopeia.

NF Grade: Meets the requirements of the National Formulary.

MB Grade (Molecular Biology): Applies to products that are suitable for a variety of Molecular Biology applications including those which require ultra-low levels of trace metal contamination, and low levels of DNase, RNase, and protease. These products are also functionally tested when applicable.

Reagent Grade: Suitable for use in general laboratory applications; meets in-house established limits in the absence of any compendial reference for the given compound. In-house established limits and their associated test methods are in many cases derived from common compendial methods such as USP, NF, ACS, etc.

High Purity Grade: Generally meets more stringent requirements than Reagent Grade products, such as higher assay limits or lower levels of trace contaminants. When available, this grade is more suitable as a reference material than the Reagent Grade.

A Few Notes About USB Biochemicals

USB® has been supplying bulk biochemicals to diagnostic, biotech, and pharmaceutical companies for over 40 years.

�� Bulk quantities available�� Lot reserve and multiple lot testing�� Long term supply agreements�� MSDS and lot-specific Certificates of Analysis (C of As)�� Scale from small to large production runs�� Complete QA and QC assays

Consistency and superior quality are essential. Turn to USB as your single sourcing solution for raw materials, and to ensure your supply meets demand.


Bulk Biochemicals

Biochemicals


Ac-Co A ............Acetyl-coenzyme A iCDH ................... Isocitrate dehydrogenaseAc-P .................Acetyl phosphate IDP ...................... Inosine-5'-diphosphateAChE ................Acetylcholinesterase IMP ..................... Inosine-5'-monophosphateADH .................Alcohol dehydrogenase INT ...................... Iodonitrotetrazolium violetADP ..................Adenosine-5'-diphosphate ITP ...................... Inosine-5'-triphosphateALD ..................Fructose-1, 6-diphosphate aldolase dITP .................... 2'-Deoxyinosine-5'-triphosphateAIDH.................Aldehyde dehydrogenase ddITP .................. 2',3'-Dideoxyinosine-5'-triphosphateAMP .................Adenonsine-5'-monophosphate LDH ..................... L-Lactate dehydrogenaseD-AOD ..............D-Amino acid oxidase D-LDH ................. D-Lactate dehydrogenaseL-AOD ..............L-Amino acid oxidase MDH ................... L-Malate dehydrogenaseAP ....................Alkaline phosphatase MK ...................... MyokinaseASOD ...............Ascorbate oxidase MPI ..................... Mannosephosphate isomeraseATP ...................Adenosine-5'-triphosphate MUTARO ............. MutarotasedATP .................2'-Deoxyadenosine-5'-triphosphate NAD+ ................. Nicotinamide-adenine dinucleotideddATP ...............2',3'-Dideoxyadenosine-5'-triphosphate NADH ................. Nicotinamide-adenine dinucleotide, reducedATPase ..............Adenosine-5'-triphosphate NADP+ ............... Nicotinamide-adenine dinucleotide phosphateBAEE ................Benzoyl-L-arginine ethyl ester NADPH ............... Nicotinamide-adenine dinucleotide phosphate,BAPNA .............N—Benzoyl-L-arginine-p-nitroanilide reducedt-Boc ................tert-butoxy carbonyl NaOAc ................ Sodium acetateBSA ..................Bovine serum albumin NBT..................... Nitro-Blue-tetrazolium saltCBZ ..................Carbonbenzoxy NPS ..................... Nitrophenyl sulphenylChE ..................Cholinesterase 2-OG(α-KG) ........ 2-OxoglutarateCK(CPK) ............Creatine kinase (α-Ketoglutarate)CoA(CoA-SH) ....Coenzyme A PALP ................... Pyridoxal-5-phosphateCP ....................Creatine phosphate PAMP .................. Pyridoxamine-5-phosphateCTP ..................Cytidine-5'-triphosphate PC ....................... Paper chromatographydCTP.................2'-Deoxycytidine-5'-triphosphate PEP ..................... Phosphoenol pyruvateddCTP...............2'-3'-Dideoxycytidine-5'-triphosphate 6-PGDH .............. 6-Phosphogluconate dehydrogenaseDAP ..................Dihydroxyacetone phosphate 6-PG ................... 6- Phosphogluconate,DNA .................Deoxyribonucleic acid (D-Gluconate-6-phosphate)DNase ..............Deoxyribonuclease PGI ...................... Phosphoglucose isomerase,DNP ..................Dinitrophenyl (Phosphohexose isomerase),DNPyr ...............Dinitropyridyl (Glucosephosphate isomerase)DTE ..................Dithioerythritol PGK .................... 3-Phosphoglycerate kinaseDTT ...................Dithiothreitol PGIuM ................ PhosphoglucomutaseEDTA ................Ethylenediaminetetraacetic acid PGM ................... Phosphoglycerate mutaseENOL ................Enolase Pi ........................ Inorganic phosphateFAD ..................Flavin-adenine dinucleotide PK ....................... Pyruvate kinaseFDH ..................Formate dehydrogenase PL-A .................... Phospholipase AFDP ..................D-Fructose-1,6-diphosphate PL-D .................... Phospholipase DFDPase .............Fructose diphosphatase POD .................... PeroxidaseFMN .................Flavin mononucleotide Ppase .................. Pyrophosphatase, inorganicF-1-P ................D-Fructose-1-phosphate Ppi ...................... Inorganic pyrophosphateF-6-P ................D-Fructose-6-phosphate PPO..................... 2,5-Diphenyl oxazoleF-6-PK ..............Fructose-6-phosphate kinase PTA ..................... PhosphotransacetylaseFUM .................Fumarase RNA .................... Ribonucleic acidGal-1-P .............Galactose-1-phosphate tRNA ................... Transfer ribonucleic acidGAP ..................Glyceraldehyde-3-phosphate RNase ................. RibonucleaseGAPDH .............D-Glyceraldehyde-3-phosphate dehydrogenase SDH .................... Sorbitol dehydrogenase,α-GDH .............α-Glycerol phosphate dehydrogenase, (Polyol dehydrogenase) (Glycerol-1-phosphate dehydrogenase), SP ....................... Sucrose phosphorylase (Glycerol-3-phosphate dehydrogenase) TALD ................... TransaldolaseGDP ..................Guanosine-5'-diphosphate TIM ..................... Triosephosphate isomeraseGK ....................Glycerokinase Tris ...................... Tris-hydroxymethyl-aminomethaneGIDH ................L-Glutamate dehydrogenase TTP (dTTP) ........... Thymidine-5'-triphosphateGMP .................Guanosine-5'-monophosphate (2'-Deoxythymidine-5'-triphosphate)GOD .................Glucose oxidase ddTTP ................. 2',3'-dideoxythymide-5'-triphosphateGOT ..................Glucamate-oxaloacetate transaminase UDP .................... Uridine-5'-diphosphateG-1-P................D-Glucose-1-phosphate UDPG .................. Uridine-5'-diphosphate glucoseG-1,6-P

2 ...........D-Glucose-1,6-diphosphate UDPgal................ Uridine-5'-diphosphate galactose

G-6-P................D-Glucose-6-phosphate UDPGP ................ Uridine-5'-diphosphate glucose pyrophosphorylaseG-6-PDH ...........Glucose-6-phosphate dehydrogenase (Glucose-1-phosphate uridylyltransferase)GPT ..................Glutamate pyruvate transaminase UMP ................... Uridine-5'-monophosphateGR ....................Glutathione reductase UTP ..................... Uridine-5'-triphosphateGSH ..................Glutathione, reduced dUTP ................... 2'-Deoxyuridine-5'-triphosphateGSSG ................Glutathione, oxidized ddUTP ................. 2',3'-Dideoxyuridine-5'-triphosphateγ-GT .................γ-Glutamyltransferase XOD .................... Xanthine oxidaseGTP ..................Guanosine-5'-triphosphate dGTP ................2'-Deoxyguanosine-5'-triphosphate DBR: “Data For Biochemical Research” Dawson, R. M. C., ddGTP ..............2',3'-Dideoxyguanosine-5'-triphosphate Elliot, D. C.,Elliot, W. H., Jones, K. M., Oxford Science Publications (1986) HK ....................Hexokinase NRC: “Specifications and Criteria for Biochemical Compounds.” National HPLC ................High performance liquid chromatography Academy of Sciences, Washington, D.C., 3rd edition (1972).

Biochemical Acronyms

Biochemicals


Standard Atomic Weights of the Elements

Atomic Number Element Symbol Weight

89. . . . . . . . . Actinium . . . . . . . . .Ac . . . . . . . . .227.027713. . . . . . . . . Aluminum. . . . . . . . . Al . . . . . . . . . . 26.981539

95. . . . . . . . . Americium . . . . . . . .Am . . . . . . . . .243.0614* 51. . . . . . . . . Antimony . . . . . . . . .Sb . . . . . . . . . .121.75 18. . . . . . . . . Argon . . . . . . . . . . . . Ar . . . . . . . . . . 39.948 33. . . . . . . . . Arsenic . . . . . . . . . . . As. . . . . . . . . . . 74.92159 85. . . . . . . . . Astatine . . . . . . . . . .At . . . . . . . . . .209.9871* 56. . . . . . . . . Barium . . . . . . . . . . .Ba . . . . . . . . . .137.327 97. . . . . . . . . Berkelium . . . . . . . . .Bk . . . . . . . . . .247.0703* 4. . . . . . . . . . Beryllium . . . . . . . . .Be . . . . . . . . . . . . 9.012182 83. . . . . . . . . Bismuth . . . . . . . . . .Bi . . . . . . . . . .208.98037 107 . . . . . . . Bohrium . . . . . . . . . .Bh. . . . . . . . . .264.12 5. . . . . . . . . . Boron . . . . . . . . . . . .B. . . . . . . . . . . . 10.811 35. . . . . . . . . Bromine . . . . . . . . . .Br . . . . . . . . . . . 79.904 48. . . . . . . . . Cadmium . . . . . . . . .Cd. . . . . . . . . .112.411 55. . . . . . . . . Cesium . . . . . . . . . . .Cs . . . . . . . . . .132.90543 20. . . . . . . . . Calcium . . . . . . . . . .Ca. . . . . . . . . . . 40.078 98. . . . . . . . . Californium. . . . . . . .Cf . . . . . . . . . .251.0796* 6. . . . . . . . . . Carbon . . . . . . . . . . .C. . . . . . . . . . . . 12.011 58. . . . . . . . . Cerium . . . . . . . . . . .Ce. . . . . . . . . .140.115 17. . . . . . . . . Chlorine . . . . . . . . . .Cl . . . . . . . . . . . 35.4527 24. . . . . . . . . Chromium. . . . . . . . .Cr . . . . . . . . . . . 51.9961 27. . . . . . . . . Cobalt . . . . . . . . . . .Co. . . . . . . . . . . 58.9332 29. . . . . . . . . Copper . . . . . . . . . . .Cu. . . . . . . . . . . 63.546 96. . . . . . . . . Curium . . . . . . . . . . .Cm . . . . . . . . .247.0704* 105 . . . . . . . Dubnium. . . . . . . . . .Db. . . . . . . . . .262.1141 66. . . . . . . . . Dysprosium. . . . . . . .Dy . . . . . . . . . .162.5 99. . . . . . . . . Einsteinium . . . . . . . .Es . . . . . . . . . .252.0830 68. . . . . . . . . Erbium . . . . . . . . . . .Er . . . . . . . . . .167.26 63. . . . . . . . . Europium . . . . . . . . .Eu . . . . . . . . . .151.965 100 . . . . . . . Fermium . . . . . . . . . .Fm . . . . . . . . .257.0951 9. . . . . . . . . . Fluorine . . . . . . . . . .F . . . . . . . . . . . . 18.9984032 87. . . . . . . . . Francium. . . . . . . . . .Fr . . . . . . . . . .223.0197 64. . . . . . . . . Gadolinium. . . . . . . .Gd. . . . . . . . . .157.25 31. . . . . . . . . Gallium . . . . . . . . . .Ga. . . . . . . . . . . 69.723 32. . . . . . . . . Germanium. . . . . . . .Ge. . . . . . . . . . . 72.61 79. . . . . . . . . Gold . . . . . . . . . . . . .Au. . . . . . . . . .196.96654 72. . . . . . . . . Hafnium . . . . . . . . . .Hf . . . . . . . . . .178.49 108 . . . . . . . Hassium . . . . . . . . . .Hs 2. . . . . . . . . . Helium . . . . . . . . . . .He. . . . . . . . . . . . 4.002602 67. . . . . . . . . Holmium . . . . . . . . .Ho. . . . . . . . . .164.93032 1. . . . . . . . . . Hydrogen . . . . . . . . .H. . . . . . . . . . . . . 1.00794 49. . . . . . . . . Indium . . . . . . . . . . . In . . . . . . . . . .114.82 53. . . . . . . . . Iodine . . . . . . . . . . . . I . . . . . . . . . . .126.90447 77. . . . . . . . . Iridium . . . . . . . . . . . Ir . . . . . . . . . . .192.22 26. . . . . . . . . Iron . . . . . . . . . . . . .Fe . . . . . . . . . . . 55.847 36. . . . . . . . . Krypton . . . . . . . . . .Kr . . . . . . . . . . . 83.8 57. . . . . . . . . Lanthanum . . . . . . . .La . . . . . . . . . .138.9055 103 . . . . . . . Lawrencium . . . . . . .Lr . . . . . . . . . .262.1097 82. . . . . . . . . Lead . . . . . . . . . . . . .Pb . . . . . . . . . .207.2 3. . . . . . . . . . Lithium . . . . . . . . . . .Li . . . . . . . . . . . . 6.941 71. . . . . . . . . Lutetium . . . . . . . . . .Lu . . . . . . . . . .174.967 12. . . . . . . . . Magnesium. . . . . . . .Mg . . . . . . . . . . 24.305 25. . . . . . . . . Manganese. . . . . . . .Mn . . . . . . . . . . 54.93805 109 . . . . . . . Meitnerium . . . . . . . .Mt. . . . . . . . . .268.1388 101 . . . . . . . Mendelevium . . . . . .Md . . . . . . . . .258.0984* 80. . . . . . . . . Mercury . . . . . . . . . .Hg. . . . . . . . . .200.59

42. . . . . . . . . Molybdenum. . . . . . .Mo . . . . . . . . . . 95.94 60. . . . . . . . . Neodymium . . . . . . .Nd . . . . . . . . .144.24

Atomic Number Element Symbol Weight

10. . . . . . . . . Neon. . . . . . . . . . . . .Ne. . . . . . . . . . . 20.1797 93. . . . . . . . . Neptunium . . . . . . . .Np. . . . . . . . . .237.0482* 28. . . . . . . . . Nickel . . . . . . . . . . . .Ni . . . . . . . . . . . 58.69 41. . . . . . . . . Niobium . . . . . . . . . .Nb. . . . . . . . . . . 92.90638 7. . . . . . . . . . Nitrogen . . . . . . . . . .N. . . . . . . . . . . . 14.00674 102 . . . . . . . Nobelium . . . . . . . . .No. . . . . . . . . .259.1010 76. . . . . . . . . Osmium . . . . . . . . . .Os . . . . . . . . . .190.2 8. . . . . . . . . . Oxygen . . . . . . . . . . .O. . . . . . . . . . . . 15.9994 46. . . . . . . . . Palladium . . . . . . . . .Pd . . . . . . . . . .106.42 15. . . . . . . . . Phosphorus . . . . . . .P . . . . . . . . . . . . 30.973762 78. . . . . . . . . Platinum . . . . . . . . . .Pt . . . . . . . . . .195.08 94. . . . . . . . . Plutonium . . . . . . . . .Pu . . . . . . . . . .244.0642* 84. . . . . . . . . Polonium. . . . . . . . . .Po . . . . . . . . .208.9824* 19. . . . . . . . . Potassium . . . . . . . . .K . . . . . . . . . . . 39.0983 59. . . . . . . . . Praseodymium . . . . .Pr . . . . . . . . . .140.90765 61. . . . . . . . . Promethium . . . . . . .Pm . . . . . . . . .144.9127* 91. . . . . . . . . Protactinium . . . . . . .Pa . . . . . . . . . .231.0359 88. . . . . . . . . Radium . . . . . . . . . .Ra . . . . . . . . . .226.0254* 86. . . . . . . . . Radon. . . . . . . . . . . .Rn. . . . . . . . . .222.0176* 75. . . . . . . . . Rhenium . . . . . . . . . . Re . . . . . . . . .186.207 45. . . . . . . . . Rhodium. . . . . . . . . .Rh. . . . . . . . . .102.9055 37. . . . . . . . . Rubidium . . . . . . . . .Rb. . . . . . . . . . . 85.4678 44. . . . . . . . . Ruthenium . . . . . . . .Ru. . . . . . . . . .101.07 104 . . . . . . . Rutherfordium. . . . . .Rf . . . . . . . . . .261.1088 62. . . . . . . . . Samarium . . . . . . . . .Sm . . . . . . . . .150.36 21. . . . . . . . . Scandium . . . . . . . . .Sc . . . . . . . . . . . 44.95591 106 . . . . . . . Seaborgium. . . . . . . .Sg . . . . . . . . . .266.1219 34. . . . . . . . . Selenium. . . . . . . . . .Se . . . . . . . . . . . 78.96 14. . . . . . . . . Silicon. . . . . . . . . . . .Si . . . . . . . . . . . 28.0855 47. . . . . . . . . Silver. . . . . . . . . . . . .Ag. . . . . . . . . .107.8682 11. . . . . . . . . Sodium . . . . . . . . . . .Na. . . . . . . . . . . 22.989768 38. . . . . . . . . Strontium . . . . . . . . .Sr . . . . . . . . . . . 87.62 16. . . . . . . . . Sulfur . . . . . . . . . . . .S . . . . . . . . . . . 32.066 73. . . . . . . . . Tantalum. . . . . . . . . .Ta . . . . . . . . . .180.9479 43. . . . . . . . . Technetium . . . . . . . .Tc . . . . . . . . . . . 97.9072* 52. . . . . . . . . Tellurium. . . . . . . . . .Te . . . . . . . . . .127.6 65. . . . . . . . . Terbium. . . . . . . . . . .Tb . . . . . . . . . .158.92534 81. . . . . . . . . Thallium . . . . . . . . . .Tl . . . . . . . . . .204.3833 90. . . . . . . . . Thorium . . . . . . . . . .Th . . . . . . . . . .232.0381* 69. . . . . . . . . Thulium. . . . . . . . . . .Tm . . . . . . . . .168.93421 50. . . . . . . . . Tin . . . . . . . . . . . . . .Sn . . . . . . . . . .118.71 22. . . . . . . . . Titanium . . . . . . . . . .Ti . . . . . . . . . . . 47.88 74. . . . . . . . . Tungsten . . . . . . . . . .W . . . . . . . . . .183.85 112 . . . . . . . Ununbium. . . . . . . . .Uub 116 . . . . . . . Ununhexium . . . . . . .Uuh 110 . . . . . . . Ununnilium . . . . . . . .Uun 114 . . . . . . . Ununquadium. . . . . .Uuq 111 . . . . . . . Unununium. . . . . . . .Uuu. . . . . . . . .272.1535 92. . . . . . . . . Uranium . . . . . . . . . .U. . . . . . . . . . .238.0508* 23. . . . . . . . . Vanadium . . . . . . . . .V . . . . . . . . . . . 50.9415 54. . . . . . . . . Xenon. . . . . . . . . . . .Xe . . . . . . . . . .131.29 70. . . . . . . . . Ytterbium . . . . . . . . .Yb. . . . . . . . . .173.04 39. . . . . . . . . Yttrium . . . . . . . . . . . Y . . . . . . . . . . . 88.90585 30. . . . . . . . . Zinc. . . . . . . . . . . . . .Zn . . . . . . . . . . . 65.39 40. . . . . . . . . Zirconium . . . . . . . . .Zr . . . . . . . . . . . 91.224*Relative atomic mass of the isotope of that element of longest known half-life.Names of individual elements with atomic numbers 110 to 116 are provisional.Based on 2001 IUPAC Table of Standard Atomic Weights of the Elements.Reproduced from Pure Appl. Chem., (2003) Vol. 75, No. 8, pp. 1107-1122 by permission of the copyright owner, IUPAC. ©2003 IUPAC.

Biochemicals


In 1966, Good, et al. introduced a series of new biological buffers for use in the physiological pH range(1). Prior to that time only a few buffer systems, consisting of weak acid/salt or weak base/salt were available to biologi-cal laboratories. Most of these systems had severe limitations associated with them and are discussed later. The “Good” zwitterionic buffers were designed to meet a genuine need, particularly in the developing field of cellular biology, and have gained rapid acceptance. Zwitterionic buffers, by definition, contain both positive and negative ionizable groups. Secondary and tertiary amines provide the positive charges, while sulfonic and carbox-ylic acid groups provide the negative charges.

There are very few traditional materials which are weak acids or weak bases with pKa values effective between pH 6.0 and 9.0 where most biological processes take place. All of these compounds exhibit some significant devia-tions from the requirements of the “ideal” buffer.

Phosphate is an active participant in many biochemical processes, being an activator, inhibitor and/or a metabolite in many biological systems. Borate buffers complex with a wide variety of important respiratory metabolites and other organic compounds as well. For example, polyols, including carbohy-drates and their derivatives, can significantly lower the pH of borate buffers through complex formation. Phosphates also demonstrate complexing capabilities with divalent and polyvalent cations and can therefore inhibit a series of metal ion-dependent biochemical reactions.

Tris (also a popular traditional biochemical buffer) is a poor buffer below pH 7.5 and is a potentially reactive primary amine which often acts as an inhibitor. It has an appreciable solubility in organic solvents which permits it to penetrate biological membranes and it has a very high pH temperature coefficient.

Bicarbonate is a poor buffer due to the limited solubility of carbon dioxide and the associated problems of liquid/gas equilibrium. Since bicarbonate buffers spontaneously liberate carbon dioxide (equation 1) they require sealed vessels capable of maintaining a carbon dioxide atmosphere in order to sustain a constant pH.

1. CO3= + 2H+ HCO3

- + H+ H2O+CO2

Bicarbonate’s pKa is 6.3 and therefore has a greatly diminished buffering capacity within the physiologically important pH range of 7 to 7.5.

From the Henderson-Hassellbach equation, (equation 2), it can be shown that optimal buffering occurs when the working pH is equal to the pKa of the ionizing group, and falls at an increasing distance from that value. In practice, the useful pH range for a buffer is its pKa ± 1 pH unit.

2. pH = pKa + log [(activity of base) ÷ (activity of acid)]

Good’s buffers were designed to meet the following criteria of an effective “ideal” biological buffer as much as possible:

� pKa between 6.0 and 8.0, the region of most biological reactions� High aqueous solubility� Minimal penetration of biological membranes� Minimal salt effects or effects due to the ionic composition of the solution� Minimal effects due to concentration or temperature� Minimal interactions with mineral cations� Enzymatic and hydrolytic stability� Minimal absorbance between 240 and 700 nm� Easy to purify� Non-toxic� Minimal participation in biochemical reactions

Helpful suggestions1. When preparing concentrated solutions refer to solubility data.2. Media that contain buffers may be adjusted to correct pH with NaOH or

HCl. If this is not feasible, the buffer stock can be adjusted to correct pH before addition to the media.

3. Solutions containing buffers can be sterilized by membrane filtration or normal autoclaving procedures.

4. Don’t autoclave solutions which have been adjusted with Sodium Bicarbonate.

5. PIPES requires the addition of alkali to effect solution.6. Always use pyrogen-free distilled water when preparing media from dry

chemicals.7. Always choose a buffer with a pKa close to the pH that is required. If

the pH is expected to drop during the project, select a buffer with a pKa slightly lower than the pH.

8. Always adjust the pH of the crystalline buffers when placed into solution, otherwise the pH will be incorrect in most instances.

9. Use tetramethylammonium hydroxide when a buffer without mineral cations is required.

10. Whenever possible prepare the buffer solution at the concentration and temperature of the experimental conditions under which it will be used.

11. Other buffers suitable for ultraviolet spectroscopy include many zwitterionic acids. For example, at concentrations of 0.05 M, BES, Bicine, HEPES, MES, PIPES,TES, and Tricine have negligible absorption above 240 nm and can be used successfully at 230 nm. ACES absorbs significantly at 230 nm and ADA is limited to above about 260 nm.

References:1. Good, N. E., Winget, G. D., Winter, W., Connolly, T. N., Izawa, S. and

Singh, R. M. M. (1966) Biochemistry 5, 467-477.2. Schwarzenbach, G. (1955) Helv. Chim. Acta. 38, 1147.3. Good, N. E. and Izawa, S. (1972) Methods Enzymol. 24, 53-68.

Good’s Buffers Guidelines

Biochemicals


∆ pKa/°C Molarity of Common Formula Temperature Saturated Name Formula Weight Coefficient Solution at 0°C pKa (4°C) pKa (20°C) pKa (37°C)

ACES H2NCOCH2NH 182.20 -0.020 0.22 7.20 6.90 6.54 CH2 CH2SO3H

ADA NH2COCH2N 190.15 -0.011 0.09 6.80 6.60 6.43 (CH2 COOH)2

BES (HOCH2CH2)2NCH2 213.25 -0.016 3.2 7.41 7.15 6.88 CH2 SO3H

Bicine (CH2CH2OH)2NCH2 163.17 -0.018 1.1 8.64 8.35 8.04 COOH

CAPS (C6H11)NH(CH2)3 221.32 -0.021 0.47 10.74 10.40 10.04 SO3H

CHES (C6H11)NH(CH2)2 207.29 -0.011 1.14 9.73 9.55 9.36 SO3H

Glycinamide NH2CH2CONHCH2 132.12 -0.028 1.1 8.85 8.40 7.92 COOH

Glycinamide•HCl H2NCH2CONH2• 110.54 -0.029 4.6 8.66 8.20 7.71 HCl

HEPES HO(CH2)2(C4H8N2) 238.30 -0.014 2.25 7.77 7.55 7.31 (CH2)2SO3H

HEPPS HO3S(CH2)3 252.33 -0.011 1.58 8.18 8.00 7.81 (C4H8N2)(CH2)2OH

MES HO3S(CH2)2 213.25 -0.011 0.65 6.33 6.15 5.96 (C4H8NO)•H2O

MOPS HO3S(CH2)3 209.26 -0.011 3.09 7.38 7.20 7.01 (C4H8NO)

PIPES HO3S(CH2)2(C4H8N2) 302.37 -0.0085 _____ 6.94 6.80 6.66 (CH2)2SO3H

TES (HOCH2)3CNHCH2 229.25 -0.020 2.6 7.82 7.50 7.16 CH2SO3H

TRICINE (CH2OH)3CNHCH2 179.17 -0.021 0.8 8.49 8.15 7.79 COOH

TRIS NH2C(CH2OH)3 121.14 -0.031 2.4 8.80 8.30 7.77

References:1. Good, N. E., Winget, G. D., Winter, W., Connolly, T. N., Izawa, S. and Singh, R. M. M. (1966) Biochemistry 5, 467-477.2. Schwarzenbach, G. (1955) Helv. Chim. Acta. 38, 1147.3. Good, N. E. and Izawa, S. (1972) Methods Enzymol. 24, 53-68.

Physical Properties of Good’s and Other Biological Buffers

Biochemicals


S-Acetyl Coenzyme A, Lithium SaltULTRAPURE

C23H35N7O17P3SLi3•3H2OFormula Weight: 881.42Product specifications:Form: White powderAssay (Total UV, pH 7.0): ≥ 90%λmax: 259 ± 1 nmεmax (pH 7): [14.6 ± 0.8] x 103

Spectral Ratios:A232/A260: 0.51 ± 0.02

A250/A260: 0.86 ± 0.04 A280/A260: 0.17 ± 0.02Storage: -20°CCAS #75520-41-1

10062 10 mg 25 mg 100 mg 500 mg

N-Acetyl-L-Cysteine

HSCH2CH(NHCOCH3)COOHFormula Weight: 163.19Product specifications:Form: White crystalline powderAssay: 98.0 - 102.0% C5H9NO3S (dry basis)Identity: By IRLoss on Drying: ≤ 1.0%Residue on Ignition: ≤ 0.5%Heavy Metals (Pb): ≤ 10 ppmCAS #616-91-1

10120 100 gm 500 gm 1 kg

Acetylated BSA

See Albumin, Bovine, Acetylated, page 198

AcrylamideULTRAPURE

MB GradeCH2=CHCONH2

Formula Weight: 71.08Product specifications:Form: White/clear crystalline powderAssay: ≥ 99.9%Identity: By IRA (410 nm, 35%, 1 cm): ≤ 0.02A (290 nm, 1%, 1 cm): ≤ 0.1A (280 nm, 7 mM, 1 cm): ≤ 0.04A (252 nm, 7 mM, 1 cm): ≤ 1.20Insoluble Matter (H2O): ≤ 0.005%Insoluble Matter (MeOH): ≤ 0.005%pH (10%, 0.1N NaCl): 5.5 - 7.0Melting Point: 84° ± 1°CAcrylic Acid: ≤ 0.001%Conductivity (35%): ≤ 5 µmhos/cmIron (Fe): ≤ 1 ppmLead (Pb): ≤ 1 ppmGel Performance: Passes testApplications:� Basic component for preparing polyacrylamide

gels� Used at a 19:1 ratio with bis-acrylamideDNA Size Separation by Acrylamide Percentage: DNA Size Range Concentration of (Base Pairs) Acrylamide (%) 100 - 1,000 3.5 80 - 500 5.0 60 - 400 8.0 40 - 200 12.0 10 - 100 20.0CAS #79-06-1

75820 100 gm 500 gm 1 kg

Acrylamide

Reagent GradeC3H5NOFormula Weight: 71.08Product specifications:Form: White, clear crystalline powderAssay: ≥ 99.95%Identity: By IRA (290 nm, 1%, 1 cm): ≤ 0.125pH (10%, H2O): 5.8 - 7.5Acrylic Acid Content: ≤ 0.001%Insoluble Matter: ≤ 0.005%CAS #79-06-1

32800 25 gm 100 gm 500 gm 1 kg

Acrylamide Gel Mixes and Solutions

Glycerol Tolerant Gel (GTG) Buffer, 20X Solution, page 219

RapidGel™ 40% Liquid Acrylamide Stock Solution, page 237

RapidGel-XL 6% DNA Sequencing Gel Solution, page 238

Stop Solution, page 242

Adenine

C5H5N5

Formula Weight: 135.13Product specifications:Form: White to off-white crystalline powderIdentity: By IRAssay: ≥ 98.0%Loss on Drying: ≤ 1.0%Residue on Ignition: ≤ 0.1%Sulfate (SO4): ≤ 0.028%Heavy Metals (Pb): ≤ 20 ppmArsenic (As): ≤ 2 ppmReference constants:λmax (pH 7, NRC): 261 nmεmax (pH 7, NRC): 13.4 x 103

ε260 (pH 7, DBR): 13.3 x 103

CAS #73-24-5

10430 100 gm 1 kg

Adenine Sulfate, Dihydrate

(C5H5N5)2•H2SO4

•2H2OFormula Weight: 404.36Product specifications:Form: White to pale-yellow crystalline powderAssay (Total UV): ≥ 97.0%Identity: By IRSpectral Ratios (pH 1.0): A250/A260: 0.76 ± 0.04 A280/A260: 0.38 ± 0.04Moisture (KF): ≤ 10%CAS #321-30-2

10450 100 gm 1 kg

Biochemicals


Adenosine

C10H13N5O4

Formula Weight: 267.24Product specifications:Form: White crystalline powderAssay (UV): ≥ 98.0%Chromatography (TLC): Homogeneousλmax: 257 nm ± 2 nmSpectral Ratios (0.1N HCl): A250/A260: 0.85 ± 0.03 A280/A260: 0.21 ± 0.02Loss on Drying: ≤ 0.5%Reference constants:λmax (pH 7, NRC): 259 nmεmax (pH 7, NRC): 15.4 x 103

ε260 (pH 6, DBR): 14.9 x 103

CAS #58-61-7

10460 100 gm

Adenosine-5’-Diphosphate, Disodium Salt, Dihydrate(ADP)

C10H13N5O10P2Na2•2H2O

Formula Weight: 507.20Product specifications:Form: White powderAssay (Total UV): 100 ± 4%Spectral Ratios: A250/A260: 0.78 ± 0.03 A280/A260: 0.16 ± 0.02Reference constants:λmax (pH 7, NRC): 259 nmεmax (pH 7, NRC): 15.4 x 103

ε260 (pH 7, DBR): 15.4 x 103

Storage: -20°C in air-tight containers.CAS #16178-48-6

10490 5 gm 10 gm 25 gm

Adenosine-5’-Monophosphate, Free Acid

C10H14N5O7PFormula Weight: 347.22 (anhydrous)* 365.24 (monohydrate)*Product specifications:Form: White crystalline powderAssay (UV, pH 7.0 or 0.01N HCl): 99 ± 3%Identity: By IRSpectral Ratios (pH 7.0): A250/A260: 0.78 ± 0.03 A280/A260: 0.16 ± 0.02 orSpectral Ratios (0.01N HCl): A250/A260: 0.84 ± 0.03 A280/A260: 0.22 ± 0.02Moisture: ≤ 6%* Note: Moisture content may vary from anhydrous to a monohydrate.

Reference constants:λmax (pH 7, NRC): 259 nmεmax (pH 7, NRC): 15.4 x 103

ε260 (pH 7, DBR): 15.3 x 103

CAS #18422-05-4

10520 10 gm 25 gm 100 gm

Adenosine-5’-Monophosphate, Free Acid, Monohydrate(AMP)

C10H14N5O7P•H2OFormula Weight: 365.24Product specifications:Form: White crystalline powderAssay (UV): ≥ 96%λmax: 257 nm ± 2 nmSpectral Ratios (0.1N HCl): A250/A260: 0.84 ± 0.03 A280/A260: 0.22 ± 0.02Loss on Drying: ≤ 6.0%Reference constants:λmax (pH 7, NRC): 259 nmεmax (pH 7, NRC): 15.4 x 103

ε260 (pH 7, DBR): 15.3 x 103

CAS #18422-05-4

10521 25 gm 100 gm 1 kg

Adenosine-5’-Monophosphate, Disodium Salt(AMP)

C10H12N5O7PNa2•nH2O

Formula Weight: 391.19•nH2OProduct specifications:Form: White powderAssay (UV, 0.01N HCl): 98 ± 3%λmax: 257 nm ± 2 nm (0.01N HCl)Spectral Ratios (0.01N HCl): A250/A260: 0.84 ± 0.03 A280/A260: 0.22 ± 0.02Heavy Metals: ≤ 10 ppmReference constants:λmax (pH 7, NRC): 259 nmεmax (pH 7, NRC): 15.4 x 103

ε260 (pH 7, DBR): 15.3 x 103

CAS #4578-31-8

10620 25 gm 100 gm

Adenosine-5’-Triphosphate, 100 mM Solution(ATP)

ULTRAPURE

C10H12N5O13P3Na4


77241 25 µmol (250 µl)

Adenosine-5’-Triphosphate, Disodium Salt(ATP)


Formula Weight: 605.19Product specifications:Form: White crystalline powderAssay (UV, pH 7 or 0.01N HCl): 100 ± 2%λmax: 259 nm ± 2 nm (pH 7.0)εmax (pH 7.0): [15.4 ± 0.3] x 103

Spectral Ratios (pH 7.0):A250/A260: 0.80 ± 0.03

A280/A260: 0.15 ± 0.02 orSpectral Ratios (0.01N HCl): A250/A260: 0.85 ± 0.05 A280/A260: 0.22 ± 0.03Moisture: ≤ 15%Reference constants:λmax (pH 7, NRC): 259 nmεmax (pH 7, NRC): 15.4 x 103

ε260 (pH 7, DBR): 15.4 x 103

Storage: -20°C, protect from moisture.CAS #51963-61-2

10585 5 gm 10 gm 25 gm 100 gm 500 gm

Biochemicals


S-Adenosyl-L-Methionine Sulfate p-Toluenesulfonate(SAMe-PTS)

C15H23N6O5S(+) n HSO4

(-) n H2SO4C7H7SO3HFormula Weight: 766.80Product specifications:Form: White to off white powderAssay (HPLC): ≥ 95% (dry basis)Moisture (KF): ≤ 2.0%Storage: 4°CCAS #97540-22-2

10601 25 mg 100 mg

Adonitol(D-Ribitol)

C5H12O5

Formula Weight: 152.15Product specifications:Form: White to off-white powderMelting Point: 103° ± 2°CLoss on Drying: ≤ 0.5%Heavy Metals (Pb): ≤ 10 ppmCAS #488-81-3

10635 25 gm 100 gm

ADP

See Adenosine-5’-Diphosphate, Disodium Salt, Dihydrate, page 189

AEBSF Hydrochloride(4-(2-Aminoethyl)-benzenesulfonyl fluoride hydrochloride)

C8H10FNO2S•HClFormula Weight: 239.69Product specifications:Form: Fine white powderAssay (HPLC): ≥ 98%Identity: By IRMelting Point: 181 - 185°CStorage: 4°CCAS #30827-99-7Applications:� Inactivation of trypsin and thrombin(1)

� Synthetic and low molecular weight inhibitor of serum kallikrein(2)

� Specific and irreversible inhibitor of serine prote-ases(3)

Notes: AEBSF irreversibly inactivates thrombin and other serine proteases (e.g. chymotrypsin, kallikrein, plasmin, proteinase K, trypsin) by sulfonylation of a functional group in the active center of the enzyme(1). AEBSF (LD50 determined from the oral dose in mouse is 2.8 gm/kg, which is higher than those for DFP and PMSF) is a substantially less toxic substitute for PMSF and DFP. It is highly water-soluble and has an acidic pH stability. AEBSF is inactivated at 37°C by 50% after 5 hours. We recommend a concentration of stock solution of 20 mM or 100 mM in water or recommended buffer at acidic pH. Stock solutions are stable for up to 2 months at -20°C and for approximately one week at 4°C. AEBSF should be added to an assay buffer just before use at a final concentration of 0.1 - 2 mM(3-5).

References:1. Walsmann, P., Richter, M. and Markwardt, F.

(1972) Acta Biol. Med. Ger. 28, 577-585.2. Markwardt, F., Drawert, J. and Walsmann, P.

(1974) Biochem Pharmacol. 23, 2247-2256.3. Taylor, J. A., Karas, J. L., Ram, M. K., Green, O. M.

and Seidel-Dugan, C. (1995) Mol. Cell. Biol. 8, 4149-4157.

4. Murphy, B. J., Rossie, S., De Jongh, K. S. and Catterall, W. A. (1993) J. Biol. Chem. 268, 27355-27362.

5. Nathan, D. F. and Lindquist, S. (1995) Mol. Cell. Biol. 15, 3917-3925.

11118 200 mg 1 gm

Agar

Product specifications:Form: Light tan granular powderIdentity: Passes testLoss on Drying: ≤ 20.0%Total Ash: ≤ 6.5%CAS #9002-18-0

10654 1 lb 5 lb 25 lb

Agar, BacteriologicalULTRAPURE

Type AProduct specifications:Form: White to cream-colored powderMoisture: ≤ 12%Ash: ≤ 6.5%Gel Strength (1.5%): 550 - 950 gm/cm2

Gel pH (1.5%) after autoclaving: 6.0 - 7.5Gel Point (1.5%): 32 - 38°CMelting Point (1.5%): 80 - 90°CApplication:� Used extensively as a gelling agent in the prepa-

ration of bacteriological culture media. Its main attributes are superior transparency, very reliable reproducibility, and absence of inhibitors capable of hindering optimal development of microorgan-isms. Extraneous matter reduced to a minimum.

CAS #9002-18-0

10906 1 lb 5 lb 25 lb

Agar, Noble AgarULTRAPURE

Product specifications:Form: Pale cream powderMoisture: ≤ 10%Ash: ≤ 1.6%Gel Strength (1.5%): ≥ 700 gm/cm2

Gel pH (1.5%): 5.0 - 7.0Gel Point (1.5%): 32 - 37.5°CMelting Point (1.5%): 80 - 95°CA (430 nm, 1.5%): ≤ 0.150Resistance (1% suspension): ≥ 20,000 OhmsEEO (-mr): ≤ 0.450 (pH 8.4)Application:� Gelling agent essentially free of impurities. Used

mainly in biochemistry and molecular biology procedures wherever increased purity is required.

Typical Agar Preparation: Plates: 15 gm/l of media; 30-35 ml per 90 mm

plate Top Agar: 7 gm/l of media Autoclave for 20 minutes and swirl gently to

evenly distribute the agar. If antibiotics are to be added, allow the agar to cool to 50°C. When pouring plates, gently swirl the medium. A Bunsen burner flame can be passed over the surface of the plates to remove any air bubbles. After the medium has hardened, invert the plates and store at 4°C.

CAS #9002-18-0

10907 100 gm 500 gm 1 kg

Biochemicals


USB Agarose Selection Guide

Genetic Performance Certified (GPC): Functionally tested in specific molecular biology applications (e.g. nucleic acid recovery, in-gel restriction enzyme digestion or ligation) to ensure optimal agarose performance.

Hi-Res: High resolution.

Biochemicals


Effect of Agarose Concentration on DNA Separation

1X TAE Buffer Gel Concentration 1X TBE Buffer

Size Range (bp) % Size Range (bp)

Agarose - Separation ≥ 500 bp, Genetic Performance Certified [75817]

20,000 - 1,000 0.6 15,000 - 1,000 12,000 - 500 0.8 10,000 - 500 8,000 - 300 1.0 7,000 - 250 6,000 - 200 1.2 5,000 - 200 3,500 - 100 1.5 3,000 - 100 2,000 - 50 2.0 2,000 - 50

Agarose - Hi-Res, Separation ≤ 1000 bp [10132]

1,500 - 100 2.0 1,200 - 100 1,000 - 50 3.0 700 - 40 500 - 20 4.0 200 - 20 300 - 10 5.0 < 100

Agarose - LE [32802]

20,000 - 1,000 0.6 15,000 - 1,000 12,000 - 500 0.8 10,000 - 500 8,000 - 300 1.0 7,000 - 250 6,000 - 200 1.2 5,000 - 200 3,500 - 100 1.5 3,000 - 100 2,000 - 50 2.0 2,000 - 50

Agarose - Low Melt, Separation ≤ 1000 bp, Genetic Performance Certified [32829]

1,500 - 500 2.0 1,000 - 400 700 - 150 3.0 500 - 100 300 - 70 4.0 150 - 10 50 - 10 5.0 < 30

Agarose - Low Melt, Separation ≥ 1000 bp, Genetic Performance Certified [32830]

20,000 - 500 0.75 12,000 - 500 16,000 - 300 1.00 8,000 - 300 10,000 - 250 1.25 4,000 - 200 5,000 - 200 1.50 3,000 - 150 2,500 - 100 1.75 2,000 - 100 1,500 - 50 2.00 1,000 - 50

Agarose - HEULTRAPURE

Product specifications:Form: Granular, free-flowing powderEEO (-mr): 0.23 - 0.26Gel Point (1.5%): 36 ± 1.5°CRemelt Point (1.5%): 88 ± 1.5°CGel Strength (1%): ≥ 750 gm/cm2

Gel Strength (1.5%): ≥ 1200 gm/cm2

Moisture: ≤ 7%Turbidity (1.5%): ≤ 40 NpResidue on Ignition: ≤ 1.0%Sulfate (SO4): ≤ 0.2%Applications:� Counter current electrophoresis� Immunodiffusion

CAS #9012-36-6

32804 100 gm 250 gm 500 gm

Agarose - MEULTRAPURE

Product specifications:Form: Granular, free-flowing powderEEO (-mr): 0.16 - 0.19Gel Point (1.5%): 36 ± 1.5°CRemelt Point (1.5%): 88 ± 1.5°CGel Strength (1%): ≥ 1000 gm/cm2


Moisture: ≤ 7%Turbidity (1.5%): ≤ 40 NpResidue on Ignition: ≤ 0.5%Sulfate (SO4): ≤ 0.2%Applications:� High resolution protein electrophoresis� Immunoelectrophoresis� Electro immunoassays such as rocket assays� Counter current electrophoresisCAS #9012-36-6

32803 25 gm 100 gm 250 gm 500 gm

Biochemicals


ULTRAPURE

MB GradeProduct specifications:Form: Granular, free-flowing powderEEO (-mr): 0.05 - 0.13Gel Point (1.5%): 36.0 ± 1.5°CRemelt Point (1.5%): 88.0 ± 1.5°CGel Strength (1%): ≥ 1,200 gm/cm2

Gel Strength (1.5%): ≥ 2,500 gm/cm2

Moisture: ≤ 7%Turbidity (1.5%): ≤ 40 NpResidue on Ignition: ≤ 0.4%Sulfate (SO4): ≤ 0.14%

Agarose - Separation ≥ 500 bp, Genetic Performance Certified™

* Range of separation depends upon the choice of buffer. Migration in TBE buffer is slower, therefore slightly lower concentrations can be used to obtain the same range of separation.

** Size in base pairs

Guide for separation ranges*Agarose – Separation ≥ 500 bp, Genetic Performance Certified (Product No. 75817) 200 400 600 800 1,000 2,000 4,000 6,000 8,000 10,000 12,000 14,000 16,000 18,000 20,000 bp**

TAE Buffer TBE Buffer

0

Preparation of an Agarose Gel (PNs 75817 & 32802):Weigh out the desired amount of agarose and place in an Erlenmeyer flask with a measured amount of electrophoresis buffer. Example: For an 0.8% gel, add 0.8 gm of agarose and 100 ml of TBE Buffer (1X), to a 200 ml flask. The larger flask ensures against the agarose boiling over. Dissolve the agarose in a boiling water bath or in a revolving-plate microwave oven. All the grains of agarose should be dissolved and the solution clear. Cool the solution to 60°C (70°C for concentrations 2% or above) and pour immediately. Allow the gel to set for one-half hour before using. Make sure to use the same electrophoresis buffer in the gel as for the running buffer.

Related products


5 L


5 L


Genetic performance tests:DNase: Passes testRNase: Passes testRestriction Enzyme Digestion: Passes testLigation: Passes testDNA Binding: Passes testBlotting: Passes testGel Background: Passes testDNA Resolution: Passes test

Separation guidelines:Range of separation in 1X TAE:

0.6%: 20,000-1,000 bp0.8%: 12,000-500 bp1.0%: 8,000-300 bp1.2%: 6,000-200 bp1.5%: 3,500-100 bp2.0%: 2,000-50 bp

Range of separation in 1X TBE:0.6%: 15,000-1,000 bp0.8%: 10,000-500 bp1.0%: 7,000-250 bp1.2%: 5,000-200 bp1.5%: 3,000-100 bp2.0%: 2,000-50 bp

Applications:� Separation of DNA or RNA fragments ≥ 500 bp.� PCR analysis.� Recovery of separated DNA or RNA fragments

from gel.� Blotting applications such as Southerns and

Northerns.CAS #9012-36-6

75817 25 gm 100 gm 250 gm 500 gm

Biochemicals




Guide for separation ranges*Agarose – LE (Product No. 32802)

200 400 600 800 1,000 2,000 4,000 6,000 8,000 10,000 12,000 14,000 16,000 18,000 20,000 bp**


0

ULTRAPURE

MB GradeProduct specifications:Form: Granular, free-flowing powderEEO (-mr): 0.05 - 0.13Gel Point (1.5%): 36 ± 1.5°CRemelt Point (1.5%): 88 ± 1.5°CGel Strength (1%): ≥ 1,200 gm/cm2

Gel Strength (1.5%): ≥ 2,500 gm/cm2

Moisture: ≤ 7%Turbidity (1.5%): ≤ 40 NpResidue on Ignition: ≤ 0.4%Sulfate (SO4): ≤ 0.14%DNase: Passes testRNase: Passes testGel Background: Passes test

Agarose - LE


0.6%: 20,000-1,000 bp0.8%: 12,000-500 bp1.0%: 8,000-300 bp1.2%: 6,000-200 bp1.5%: 3,500-100 bp2.0%: 2,000-50 bp

Range of separation in 1X TBE:0.6%: 15,000-1,000 bp0.8%: 10,000-500 bp1.0%: 7,000-250 bp1.2%: 5,000-200 bp1.5%: 3,000-100 bp2.0%: 2,000-50 bp

Applications:� Routine separation of DNA or RNA fragments.� PCR analysis.� Blotting applications such as Southerns and

Northerns.� Protein analysis such as immunodiffusion, rocket

assays, and immuno electrophoresis.CAS #9012-36-6

32802 25 gm 100 gm 250 gm 500 gm 1 kg

Biochemicals




Guide for separation ranges*Agarose – Hi-Res, Separation ≤ 1,000 bp (Product No. 10132) 10 20 30 40 50 60 70 80 90 100 200 300 400 500 600 700 800 900 1,000 1,100 1,200 1,300 1,400 1,500 bp**


0

ULTRAPURE

MB GradeProduct specifications:Form: Granular, free-flowing powderEEO (-mr): ≤ 0.12Gel Point (3%): ≤ 35.5°CRemelt Point (3%): ≤ 80°CGel Strength (1.5%): ≥ 600 gm/cm2

Turbidity (1.5%): ≤ 50 NpGel Strength (3%): ≥ 1,500 gm/cm2

Moisture: ≤ 7%Residue on Ignition: ≤ 0.35%

Agarose - Hi-Res, Separation ≤ 1000 bp

Sulfate (SO4): ≤ 0.11%DNase: Passes testRNase: Passes testSeparation guidelines:Range of separation in 1X TAE: 1.8%: 1,000-500 bp 2.0%: 1,500-100 bp 3.0%: 600-150 bp 4.0%: 500-20 bp 4.5%: 350-50 bp 5.0%: 300-10 bp

Range of separation in 1X TBE:2.0%: 1,200-100 bp3.0%: 700-40 bp4.0%: 200-20 bp5.0%: < 100 bp

Applications:� Highly resolved separation of DNA or RNA frag-

ments ≤ 1000 bp� PCR analysisCAS #9012-36-6

10132 25 gm 100 gm 500 gm

Related products – Buffers & stock biochemicals:

Ethidium Bromide32813 1 gm 5 gm 25 gm





Related products – DNA markers:

PCR Markers, 50 – 2000 bp76710 250 µl

DNA Ladder, 100 bp76712 500 µl


Biochemicals


ULTRAPURE

MB GradeProduct specifications:Form: Granular, free-flowing powderEEO (-mr): ≤ 0.12Gel Point (1.5%): 24 - 28°CRemelt Point (1.5%): ≤ 65.5°CGel Strength (1%): ≥ 250 gm/cm2


Moisture: ≤ 7%Turbidity (1.5%): ≤ 40 NpResidue on Ignition: ≤ 0.4%Sulfate (SO4): ≤ 0.12%

Agarose - Low Melt Separation ≥ 1000 bp, Genetic Performance Certified

Genetic performance tests:DNase: Passes testRNase: Passes testIn-Gel Restriction Enzyme Digestion: Passes testIn-Gel Ligation: Passes testDNA Binding: Passes testGel Background: Passes testDNA Resolution: Passes testAgarase Digestion: Passes test


0.75%: 20,000 - 500 bp1.00%: 16,000 - 300 bp1.25%: 10,000 - 250 bp1.50%: 5,000 - 200 bp1.75%: 2,500 - 100 bp2.00%: 1,500 - 50 bp

Range of separation in 1X TBE:0.75%: 12,000 - 500 bp1.00%: 8,000 - 300 bp1.25%: 4,000 - 200 bp1.50%: 3,000 - 150 bp1.75%: 2,000 - 100 bp2.00%: 1,000 - 50 bp

Applications:� Enzymatic modification of nucleic acids in re-

melted gels.� Separation of DNA or RNA fragments ≥ 1,000 bp.� In-gel recovery of nucleic acid fragments.� PCR analysis.� Labile sample procedures.� Suitable for cell culture and cloning applications.CAS #9012-36-6

32830 25 gm 100 gm



Guide for separation ranges*Agarose – Low Melt Separation ≥ 1,000 bp, Genetic Performance Certified (Product No. 32830)

Isolating DNA from low-melting temperature agarose (PNs 32829 & 32830):Follow procedure for preparing an agarose gel and be sure to run the electrophoresis at 4°C to keep the gel from melting. Excise the DNA band of interest and transfer it to a microcentrifuge tube. Cut as close to the DNA band as possible to avoid any unnecessary agarose. Add 5 volumes of 20 mM Tris-HCl (pH 8.0), 1 mM EDTA (pH 8.0) to the tube with the agarose. Cap the tube and heat for 5 minutes at 65°C. The aga-rose should melt. Cool the solution to room temperature and add an equal volume of equilibrated phenol. Vortex for 20 seconds and spin at 400 x g for 10 minutes (20°C). Remove the aqueous phase (top) to a new microcentrifuge tube. Re-extract with phenol:chloroform and then with chloroform. Precipitate the final aqueous phase with 0.2 volumes of 10 mM ammonium acetate and 2 volumes of ethanol. Recover the DNA by centrifugation.

100 200 300 400 500 600 700 800 900 1,000 2,000 4,000 6,000 8,000 10,000 12,000 14,000 16,000 18,000 20,000 bp**0


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Biochemicals


ULTRAPURE

MB GradeProduct specifications:Form: Granular, free-flowing powderEEO (-mr): ≤ 0.12Gel Point (4%): ≤ 35°CRemelt Point (4%): ≤ 65°CGel Strength (4%): ≥ 1,000 gm/cm2

Moisture: ≤ 5%Residue on Ignition: ≤ 0.3%Sulfate (SO4): ≤ 0.12%

Agarose - Low Melt Separation ≤ 1000 bp, Genetic Performance Certified

Genetic performance tests:DNase: Passes testRNase: Passes testIn-Gel Restriction Enzyme Digestion: Passes testIn-Gel Ligation: Passes testDNA Binding: Passes testGel Background: Passes testDNA Resolution: Passes testAgarase Digestion: Passes test


2.0%: 1,500-100 bp3.0%: 700-150 bp4.0%: 300-70 bp5.0%: 50-10 bp

Range of separation in 1X TBE:2.0%: 1,000-400 bp3.0%: 500-100 bp4.0%: 150-10 bp5.0%: < 30 bp

Applications:� Enzymatic modification of nucleic acids in re-

melted gels.� Separation of DNA or RNA fragments ≤ 1,000 bp.� In-gel recovery of nucleic acid fragments.� PCR analysis.� Labile sample procedures.CAS #9012-36-6

32829 25 gm 100 gm



Guide for separation ranges*Agarose – Low Melt Separation ≤ 1,000 bp, Genetic Performance Certified (Product No. 32829) 10 20 30 40 50 60 70 80 90 100 200 300 400 500 600 700 800 900 1,000 1,100 1,200 1,300 1,400 1,500 bp**


0

ULTRAPURE

MB GradeProduct specifications:Form: Granular, free-flowing powderEEO (-mr): ≤ 0.12Gel Point (1.5%): ≤ 20°CRemelt Point (1.5%): ≤ 63°C

Agarose - Ultra Low Melt


Moisture: ≤ 7%Turbidity (1.5%): ≤ 40 NpResidue on Ignition: ≤ 0.35%Sulfate (SO4): ≤ 0.14%DNase: Passes testRNase: Passes test

Applications:� Labile sample procedures.� Cell culture and cloning applications (hybrid-

omas).CAS #9012-36-6

32821 10 gm 25 gm

Biochemicals


β-Alanine

NH2CH2CH2COOHFormula Weight: 89.09Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (as is)Identity: By IRChromatography (TLC): HomogeneousLoss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.2%Ammonia (NH4): ≤ 0.1%Chloride (Cl): ≤ 0.04%Sulfate (SO4): ≤ 0.05%CAS #107-95-9

10665 200 gm 1 kg 5 kg

Albumin, Bovine, Protease-Free

MB GradeProtease FreeSource: Bovine BloodProduct specifications:Form: Lyophilized powderAssay (electrophoresis): ≥ 98.0%Protein: ≥ 96.0%pH (1%, 0.15 M NaCl): 6.8 - 7.2Moisture: ≤ 5.0%Residue on Ignition: ≤ 1.5%IgG: Passes testHeavy Metals (Pb): ≤ 10 ppmProtease: Passes testStorage: 4°CCAS #9048-46-8

10867 10 gm 50 gm 100 gm

Tired of columns? Try ExoSAP-IT reagent, our enzymatic PCR cleanup solution.

Albumin, Bovine, Acetylated(Acetylated BSA)

ULTRAPURE

MB GradeSource: Bovine BloodProduct specifications:Form: Clear, colorless solution in deionized water;

packed under aseptic, nuclease-free conditions.Assay: Single band by SDS-polyacrylamide gel elec-

trophoresisProtein Concentration: 25 - 50 mg/mlContaminant Testing:

DNase (endo): Passes testDNase (exo): Passes testRNase: Passes testProtease: Passes test

Functional Test: No inhibition by albumin on diges-tion of lambda DNA.

Form: Many nucleases are irreversibly inactivated by acetylation, but the stabilizing action of BSA is unaffected by this procedure.(1,2) Thus, acetylation ensures that the use of albumin does not intro-duce additional nucleases to reaction mixture.

Solubility: Soluble in waterApplication:� Bovine Serum Albumin is used extensively by

molecular biologists to stabilize labile enzymes such as restriction endonucleases, polymerases, and ligases.

References:1. Epstein, C. J. and Goldberger, R. F. (1964) J. Biol.

Chem. 239, 1087-1089.2. Lackey, D. and Linn, S. (1980) Meth. in Enzymol.,

65, 26-28.Storage: -20°CCAS #9048-46-8

10848 25 mg 125 mg

Albumin, Bovine, Crystallized, pH 5.2

Source: Bovine BloodProduct specifications:Form: Crystallized powderAssay (electrophoresis): ≥ 99%Identity: By IRProtein: ≥ 98%pH (7%, 0.15 M NaCl): 5.0 - 5.4Moisture (KF) when packaged: ≤ 5.0%Sodium (Na): 0 - 5.0 mg/gmChloride (Cl): 0 - 3.0 mg/gmStorage: -20°CCAS #9048-46-8

10856 5 gm 10 gm 100 gm

Albumin, Bovine, Cohn Fraction V, pH 5.2(Cold Alcohol Fractionated)

Source: Bovine BloodProduct specifications:Form: Lyophilized powderAssay (electrophoresis): ≥ 96%Protein: ≥ 95%pH (1%, H2O): 5.0 - 5.4Residue on Ignition: ≤ 1.5%Loss on Drying: ≤ 6.0%Heavy Metals (Pb): ≤ 50 ppmStorage: 4°C, desiccatedCAS #9048-46-8

10852 50 gm 100 gm 500 gm

Albumin, Bovine, pH 7(Heat-Shock Fractionated)

Source: Bovine BloodProduct specifications:Form: Lyophilized powderAssay (electrophoresis): ≥ 98%Protein: ≥ 96.0%pH (2%, H2O): 6.5 - 7.5Loss on Drying: ≤ 5.0%Residue on Ignition: ≤ 2.0%Storage: 4°CCAS #9048-46-8

10857 50 gm 100 gm 500 gm 1 kg

Albumin, Bovine, RIA Grade, pH 7

Source: Bovine BloodProduct specifications:Form: Lyophilized powderAssay (electrophoresis): ≥ 98.0%Protein: ≥ 96.0%pH (1%. 0.15 M NaCl): 6.8 - 7.2Loss on Drying: ≤ 5.0%Residue on Ignition: ≤ 2.0%IgG: NegativeInsulin: NegligibleFolic Acid: NegligibleT3, T4: NegligibleApplication:� Suitable for use in radioimmuno assayStorage: 4°CCAS #9048-46-8

10868 10 gm 50 gm 100 gm

Biochemicals


Albumin, Egg(Ovalbumin)

Source: Egg White, Spray DriedProduct specifications:Form: Off-white crystalsProtein (as is): ≥ 80%pH: 6.5 - 8.0Moisture: ≤ 8.0%Salmonella: NegativeCAS #9006-59-1

15460 1 lb 5 lb 25 lb

Albumin, Egg(Ovalbumin)

Source: Chicken EggProduct specifications:Form: White to pale yellow freeze dried powderAssay: ≥ 98.0% (as confirmed by SDS-PAGE)Moisture: ≤ 6.0%Solubility: Soluble in distilled water or dilute bufferStorage: -20°CCAS #9006-59-1

10865 5 gm

Alcohol Dehydrogenase

(E.C.1.1.1.1)Source: YeastProduct specifications:Form: Lyophilized powderActivity: ≥ 300 units/mg proteinUnit Definition: One unit reduces 1 µmol NAD/min-

ute at 25°C, pH 8.8.Contaminating Activities:Carboxylase: ≤ 0.05%GAPDH: ≤ 0.05%PGK: ≤ 0.05%MK: ≤ 0.03%Aldolase: ≤ 0.02%PK: ≤ 0.01%LDH: ≤ 0.01%Storage: -20°CCAS #9031-72-5

10895 75,000 units

Aminoacetic Acid

See Glycine, page 219

4-Amino Antipyrene

C11H13N3OFormula Weight: 203.24Product specifications:Form: Pale yellow to light amber crystalline powderAssay: ≥ 98.0%Identity: By IRSolubility (5%, H2O): Clear solutionMelting Point: 109 ± 2°CCAS #83-07-8

11056 50 gm 100 gm

p-Amino Benzoic Acid (PABA)

USBioAnalyzedC7H7NO2

Formula Weight: 137.14Product specifications:Form: White to off-white crystalline powderAssay (dry basis): 98.5 - 101.5%Identity (A, B): Passes testsMelting Point: 186 - 189°CLoss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 0.002%Volatile Diazotizable Substances: ≤ 0.002%Ordinary Impurities: Passes testCAS #150-13-0

11055 200 gm 500 gm 1 kg 5 kg

L-α-Amino-N-Butyric Acid

CH3CH2CH(NH2)COOHFormula Weight: 103.12Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (as is)Identity: By IRChromatography (TLC): HomogeneousMoisture (KF): ≤ 1%CAS #1492-24-6

11079 1 gm 5 gm

2-Aminoethane Sulfonic Acid

See Taurine, page 243

L-α-Aminoglutaric Acid

See L-Glutamic Acid, page 218

6-Amino Purine

See Adenine, page 188

Ammonium AcetateULTRAPURE

ACS Reagent GradeCH3COONH4

Formula Weight: 77.08Product specifications:Form: White/clear crystalsAssay: ≥ 97.0%pH (5%, H2O): 6.7 - 7.3Insoluble Matter: ≤ 0.005%Residue on Ignition: ≤ 0.01%Chloride (Cl): ≤ 5 ppmNitrate (NO3): ≤ 0.001%Sulfate (SO4): ≤ 0.001%Heavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 5 ppmSolubility (5 M, H2O): Clear, colorlessApplications:� Used in the precipitation of DNA� Said to help decrease the co-precipitation of

dNTPs� Asymmetric PCR precipitationStock Solution, 7.5 M: Ammonium Acetate: 289 gm dH2O:175 ml Adjust volume with water to 500 ml. Filter sterilize.CAS #631-61-8

11251 500 gm 1 kg

Ammonium Acetate (NH4OAc), 5 M SolutionULTRAPURE

MB GradeForm: 5 M solution of ammonium acetate (NH4OAc)

in high purity dH2O. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles.

Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test

75901 100 ml 500 ml

Biochemicals


Ammonium Persulfate (APS)(Ammonium Peroxydisulfate)

ULTRAPURE

ACS Reagent Grade(NH4)2S2O8

Formula Weight: 228.19Product specifications:Form: Off-white crystalline powderAssay: ≥ 98.0%Residue on Ignition: ≤ 0.05%Insoluble Matter: ≤ 0.005%Titratable Free Acid: ≤ 0.04 meq/gmChloride and Chlorate (Cl): ≤ 0.001%Heavy Metals (Pb): ≤ 0.005%Iron (Fe): ≤ 0.001%Manganese (Mn): ≤ 0.5 ppmApplication:� A high speed initiator used to catalyze acrylamide

polymerizationWorking Stock Solution (10%): Ammonium Persulfate: 1 gm dH2O: to 10 ml This solution should be stored at 4°C and pre-

pared fresh weekly.CAS #7727-54-0

76322 10 gm 100 gm 1 kg

Ammonium SulfateULTRAPURE

ACS Reagent Grade(NH4)2SO4

Formula Weight: 132.14Product specifications:Form: White crystalline powderAssay: ≥ 99.0%pH (5%, H2O, 25°C): 5.0 - 6.0Insoluble Matter: ≤ 0.005%Residue on Ignition: ≤ 0.005%Chloride (Cl): ≤ 5 ppmNitrate (NO3): ≤ 0.001%Phosphate (PO4): ≤ 5 ppmHeavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 5 ppmApplication:� Used in the purification of enzymes and other

proteins by precipitationCAS #7783-20-2

11254 1 kg 5 kg

AMP

See Adenosine-5’-Monophosphate, Disodium Salt, page 189

Ampicillin, Sodium SaltULTRAPURE

USBioAnalyzedC16H18N3NaO4SFormula Weight: 371.39Product specifications:Form: White to off-white powderActivity: 845 µg - 988 µg of C16H19N3O4S/mg (dry

basis)Activity: 828 µg - 988 µg of C16H19N3O4S/mg (as is)Identity (A, B): Passes testspH (10 mg/ml): 8.0 - 10.0Moisture (KF): ≤ 2.0%Dimethylaniline: Meets requirementsMethylene chloride: ≤ 0.2%Storage: Controlled room temperature, in tightly

closed containers.CAS #69-52-3

11259 5 gm 25 gm 100 gm

Antibiotic G-418 Sulfate

See G-418 Sulfate, page 217

AprotininULTRAPURE

Source: Bovine LungC284H432N84O79S7

Formula Weight: 6511.44Product specifications:Form: Lyophilized powderActivity: 5,100 - 7,300 Kallikrein Inhibitor units/mg

proteinPurity (gel filtration): ≥ 95%Loss on Drying: ≤ 6.0%Applications:� A useful protease inhibitor against kallikrein,

trypsin, plasmin and chymotrypsin� Not effective against papainWorking Concentration: 1-2 mg/ml Soluble in 0.01 M HEPES (pH 8.0)Note: Avoid repeated freezing and thawing of stock

solutions.Storage: 4°C in air-tight containers.CAS #9087-70-1

11388 10 mg 25 mg 100 mg

L- (+)-Arabinose

C5H10O5

Formula Weight: 150.13Product specifications:Form: White to off-white crystalline powderAssay (GC): ≥ 96%Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.5%Chloride (Cl): ≤ 100 ppmSulfate (SO4): ≤ 100 ppmArsenic (As): ≤ 3 ppmIron (Fe): ≤ 50 ppmHeavy Metals (Cu): ≤ 5 ppmCAS #5328-37-0

11405 25 gm 100 gm 1 kg

L-Arabinose

High Purity GradeC5H10O5

Formula Weight: 150.13Product specifications:Form: White crystalline powder[α]20

D : +103.0° ± 1.5° (c=4 in H2O, NH3)Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Arsenic (As): ≤ 0.5 ppmIron (Fe): ≤ 5 ppmHeavy Metals (Pb): ≤ 10 ppmCAS #5328-37-0

11406 25 gm 100 gm 1 kg

L-Arginine

C6H14N4O2

Formula Weight: 174.20Product specifications:Form: White crystalline powderAssay: ≥ 98.5%Identity: By IR[α]20

D : +26.9° to +27.9° (c=8 in 6N HCl)Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.3%Heavy Metals (Pb): ≤ 15 ppmAmmonium (NH4): ≤ 0.02%Arsenic (As2O3): ≤ 1 ppmChloride (Cl): ≤ 0.05%Iron (Fe): ≤ 30 ppmSulfate (SO4): ≤ 0.03%CAS #74-79-3

11490 100 gm 500 gm 1 kg

Biochemicals


L-Arginine, Hydrochloride

NH2C(NH)NH(CH2)3CH(NH2)COOH•HClFormula Weight: 210.66Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5%Chromatography (TLC): HomogeneousIdentity: By IR[α]20

D : +21.4° to +23.6° (c=8 in 6N HCl)Loss on Drying: ≤ 0.20%Residue on Ignition: ≤ 0.10%Chloride (Cl): 16.5 to 17.1%Sulfate (SO4): ≤ 0.03%Heavy Metals (Pb): ≤ 20 ppmOrganic Volatile Impurities: Meets requirementsCAS #1119-34-2

11500 100 gm 500 gm 1 kg

L-Ascorbic Acid(Vitamin C)

USBioAnalyzedC6H8O6

Formula Weight: 176.12Product specifications:Form: White to slightly yellow crystalline powderAssay: 99.0 - 100.5%Identity A: By KBrIdentity B: Passes test[α]25

D : +20.5° to +21.5° (c=10 in H2O)Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 0.002%CAS #50-81-7

11545 1 kg 5 kg

L-Asparagine, Anhydrous

NH2COCH2CH(NH2)COOHFormula Weight: 132.12Product specifications:Form: White crystalline powderAssay: ≥ 98.5%Chromatography (TLC): HomogeneousIdentity: By IR[α]20

D : +33° to +36° (c=2 in 10% HCl)Loss on Drying: ≤ 1.0%CAS #70-47-3

11590 100 gm 500 gm 1 kg

L-Asparagine, Monohydrate

C4H8N2O3•H2O

Formula Weight: 150.13Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (as is)Identity: By IRChromatography (TLC): Homogeneous[α]20

D : +33° to +36° (c=10 in 3N HCl)Loss on Drying: ≤ 12.5%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 10 ppmArsenic (As): ≤ 3 ppmIron (Fe): ≤ 20 ppmSulfate (SO4): ≤ 0.025%Lead (Pb): ≤ 5 ppmCAS #5794-13-8

11595 100 gm 1 kg 5 kg

L-Aspartic Acid

C4H7NO4

Formula Weight: 133.10Product specifications:Form: White crystalline powderAssay: ≥ 98.0%Identity: By IRChromatography (TLC): Homogeneous[α]20

D : +24.5° to +26.0° (c=8 in 6N HCl)Loss on Drying: ≤ 0.25%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 10 ppmArsenic (As): ≤ 3 ppmCAS #56-84-8

11620 100 gm 1 kg 5 kg

L-Aspartic Acid, Sodium Salt, Monohydrate(Sodium Aspartate)

NH2CH(COOH)CH2COONa•H2OFormula Weight: 173.10Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (dry basis)Identity: By IRChromatography (TLC): Homogeneous[α]20

D : +18° to +21° (c=8 in 6N HCl, dry basis)Loss on Drying: ≤ 0.5%Heavy Metals (Pb): ≤ 10 ppmArsenic (As): ≤ 1 ppmChloride (Cl): ≤ 0.05%Iron (Fe): ≤ 30 ppmSulfate (SO4): ≤ 0.05%CAS #323194-76-9

11635 1 kg

L-Aspartic Acid, Magnesium Salt, Dihydrate

(C4H6NO4)2Mg•2H2OFormula Weight: 324.53 (dihydrate)Product specifications:Form: White powderAssay: ≥ 95.0% (dry basis)[α]20

D : +18.0° to +23.0° (20°C, c=2 in 18% HCl)Moisture (KF): ≤ 17%Heavy Metals (Pb): ≤ 10 ppmArsenic (As2O3): ≤ 1 ppmChloride (Cl): ≤ 0.021%Sulfate (SO4): ≤ 0.050%Iron (Fe): ≤ 30 ppmCAS #2068-80-6

11631 100 gm 1 kg

ATP

See Adenosine-5’-Triphosphate, page 189

Avidin-Alkaline Phosphatase Conjugate

See Streptavidin-Alkaline Phosphatase Conjugate, page 242

Biochemicals


Bacitracin

USBioAnalyzedC66H103N17O16SFormula Weight: 1422.69Product specifications:Form: White to pale buff powderActivity: ≥ 40 units of bacitracin/mg (as is)Identity: Passes testpH (10,000 units/ml): 5.5 - 7.5Loss on Drying: ≤ 5.0%Residue on Ignition: ≤ 0.5%CAS #1405-87-4

11805 5 gm 25 gm

Bacterial Culture Media

LB Agar, Ready-Made Powder, page 224LB Broth, Ready-Made Powder, page 224Luria Agar, Ready-Made Powder, page 225Luria Broth, Ready-Made Powder, page 225NZCYM Broth, Ready-Made Powder, page 231Terrific Broth, Ready-Made Powder, page 2452X YT Broth, page 251

Bacteriological Peptone(Peptones)

ULTRAPURE

Product specifications:Form: PowderMoisture: ≤ 5.0%Ash: 4.0 - 6.65%pH (6% solution): 6.6 - 7.8Amino Nitrogen: ≥ 5.1%Total Nitrogen: ≥ 12%Protein (N x 6.38): ≥ 76%An extremely high grade of peptone prepared spe-

cifically for general bacteriological purposes.CAS #73049-73-7

20048 1 lb 5 lb 25 lb

BCIP

See 5-Bromo-4-Chloro-3-Indolyl Phosphate, page 203

Betaine, 5 M SolutionULTRAPURE

MB GradeForm: 5 M solution of betaine in deionized dH2O.Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testFunctional Test: Functionally tested for use in PCR.Storage: -20°C

77507 1.5 ml 5 x 1.5 ml 10 ml

Bicine(N,N-bis[2-Hydroxyethyl]glycine)

(CH2CH2OH)2NCH2COOHFormula Weight: 163.17Product specifications:Form: White/clear crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity: By IRpKa (20°C): 8.35 ± 0.15A (250 nm, 10%, 1 cm): ≤ 0.1Loss on Drying: ≤ 0.5%Iron (Fe): ≤ 30 ppmLead (Pb): ≤ 10 ppmCAS #150-25-4

12091 100 gm 1 kg

Bilirubin

C33H36N4O6

Formula Weight: 584.66Product specifications:Form: Bright orange fine powderAssay: ≥ 97.5% (By UV, Emax = 61.2 x 103, as is)Identity: By IRMoisture (KF): ≤ 2.0%λmax: 453 ± 2 nmStorage: Store protected from light.CAS #635-65-4

12110 1 gm 5 gm

Bis-Acrylamide

See N, N’-Methylene-bis-Acrylamide, page 228

Bis-Tris(bis[2-Hydroxyethyl]imino-tris[hydroxymethyl]methane)

ULTRAPURE

(HOCH2CH2)2NC(CH2OH)3

Formula Weight: 209.24Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (20°C): 6.50 ± 0.15A (280 nm, 1 M, 1 cm): ≤ 0.4Loss on Drying: ≤ 0.5%Iron (Fe): ≤ 30 ppmLead (Pb): ≤ 10 ppmCAS #6976-37-0

12112 100 gm 500 gm 1 kg

Blotting Reagents

Denhardt’s Solution, 50X, page 209SSC, 20X Solution, page 241SSPE, 20X Solution, page 241TBS, 20X Solution, pH 7.4, page 244TBS with Tween (TBST), 20X Solution, pH 7.4,

page 244

Boric AcidULTRAPURE

MB GradeH3BO3

Formula Weight: 61.83Product specifications:Form: White crystalline powderAssay: ≥ 99.5%Insoluble in MeOH: ≤ 0.005%Nonvolatile with MeOH: ≤ 0.05%Sulfate (SO4): ≤ 0.01%Heavy Metals (Pb): ≤ 0.001%Arsenic (As): ≤ 1 ppmCalcium (Ca): ≤ 0.005%Chloride (Cl): ≤ 0.001%Iron (Fe): ≤ 0.001%Phosphate (PO4): ≤ 0.001%Functional Test: Meets or exceeds criteria for high

quality sequencing gel electrophoresis with Sequenase T7 DNA Polymerase.

DNase (endo): Passes testDNase (exo): Passes testRNase: Passes testProtease: Passes testCAS #10043-35-3

76324 500 gm 1 kg

Biochemicals


Brewers Yeast

See Yeast, Brewers, page 251

Brilliant Blue G(Coomassie® Brilliant Blue G-250)

ULTRAPURE

(C.I.42655)C47H48N3S2O7NaFormula Weight: 854.02Product specifications:Form: Dark blue/purple crystalline powderDye content: Approx. 60%Loss on Drying: ≤ 8%λmax: 607 nm ± 1 nmεmax: 38 x 103

Application:� Used for quantitative staining of proteins on poly-

acrylamide electrophoresis gels.CAS #6104-58-1

32812 25 gm 100 gm

Brilliant Blue R(Coomassie Brilliant Blue R-250)

ULTRAPURE

(C.I.42660)C45H44N3O7S2NaFormula Weight: 825.97Product specifications:Form: Dark purple/violet powderIdentity: By IRλmax: 586 ± 2 nmεmax: ≥ 45.0 x 103

Moisture (KF): ≤ 6.0%Application:� Used for quantitative staining of proteins on poly-

acrylamide electrophoresis gels.CAS #6104-59-2

32826 25 gm 100 gm

5-Bromo-4-Chloro-3-Indolyl-α-D-Galactopyranoside(X-α-GAL)

ULTRAPURE

C14H15BrClNO6

Formula Weight: 408.63Product specifications:Form: White to off-white crystalline powderAssay (TLC): ≥ 99%Chromatography (TLC): Homogeneous Identity: By IRSolubility (1%, MeOH): Clear and colorless[α]25

D : +145.0 ± 3.0° (c=0.5, DMF:H2O 1:1)Moisture (KF): ≤ 1.0%Storage: -20°C, protect from moisture.CAS #107021-38-5

26200 100 mg 500 mg

5-Bromo-4-Chloro-3-Indolyl-β-D-Galactopyranoside(X-GAL)

ULTRAPURE

C14H15BrClNO6

Formula Weight: 408.63Product specifications:Form: White crystalline powderAssay (HPLC): ≥ 99%Chromatography (TLC): HomogeneousIdentity: By IRSolubility (2%, DMF): Clear and colorlessApplications:� A histochemical substrate for β-Galactosidase� A substrate for blue/white colony screening.

Effective concentration is approximately 50 µg/ml.

Stock Concentration: 20 mg/ml in dimethylfor-mamide in a glass or polypropylene tube.


10077 100 mg 250 mg 1 gm 2 gm 5 gm

5-Bromo-4-Chloro-3-5-Bromo-4-Chloro-3-Indolyl Phosphate, Disodium Salt(BCIP)

ULTRAPURE

C8H4BrClNNa2O4PFormula Weight: 370.43Product specifications:Form: White to off-white crystalline powderIdentity: By IRChromatography (TLC): HomogeneousStorage: -20°C, protect from moisture.CAS #102185-33-1

12387 100 mg 500 mg 1 gm

5-Bromo-4-Chloro-3-Indolyl Bromocresol Green, Sodium SaltULTRAPURE

ACS Reagent GradeC21H13Br4O5SNaFormula Weight: 720.00Product specifications:Form: Brown to orange powderClarity of Solution (0.1%, H2O): Passes testVisual Transition Interval: pH 3.8 - Yellow pH 5.4 - BlueApplication: � Used as a tracking dye in alkaline agarose gel

electrophoresis.Loading Buffer (6X): 6 mM EDTA 300 mM NaOH 18% Ficoll (in water) 0.15% Bromocresol Green 0.25% Xylene CyanolCAS #62625-32-5

12355 5 gm 25 gm

Biochemicals


Bromophenol Blue, Sodium SaltULTRAPURE

ACS Reagent GradeC19H9Br4NaO5SFormula Weight: 691.94Product specifications:Form: PowderClarity of Solution: Passes testVisual Transition Interval: pH 3.0 - Yellow pH 4.6 - BlueApplications:� Used as a tracking dye in agarose and acrylamide

gel electrophoresis.� Used in the stop solution in DNA sequencing kits.CAS #62625-28-9

12370 5 gm 10 gm

BSA

See Albumin, Bovine, page 198

Buffers


HEPES, 1 M Solution, pH 7.3, page 222PBS, 10X Solution, pH 7.4, page 232RapidRun™ Agarose Buffer, 20X Solution,

page 238TAE Buffer, 10X Solution, page 243TAE Buffer, 50X Solution, page 243TBE Buffer, 5X Solution, page 243TBE Buffer, 10X Ready-Mixed Powder, page 243TE Buffer, 1X Solution, page 244TE Buffer, 1X Solution, pH 7.0, page 244TE Buffer, 1X Solution, pH 7.0, Low EDTA, page 244TE Buffer, 1X Solution, pH 8.0, page 244TE Buffer, 1X Solution, pH 8.0, Low EDTA, page 244TE Buffer, 50X Solution, page 244Tris-Glycine (TG) Buffer, 10X Solution, page 247Tris-Glycine-SDS (TGS) Buffer,10X Solution,

page 247Tris/HCl, 1 M Solution, pH 7.0, page 248Tris/HCl, 1 M Solution, pH 7.5, page 248Tris/HCl, 1 M Solution, pH 8.0, page 248

Cacodylic Acid, Sodium Salt Trihydrate(Dimethylarsenic Acid, Sodium Salt; Sodium Cacodylic)

(CH3)2AsO2Na•3H2OFormula Weight: 214.03 (trihydrate)Product specifications:Form: White powderAssay: ≥ 98.5% (dry basis)Solubility (10%, H2O): Clear and colorlessChloride (Cl): ≤ 0.1%Sulfate (SO4): ≤ 0.3%Application: � An integral component of terminal deoxynucleo-

tidyl transferase bufferNote: Material is hygroscopicCAS #124-65-2

12488 100 gm 500 gm

Caffeine, Anhydrous(1,3,7-Trimethylxanthine)

ULTRAPURE

C8H10N4O2

Formula Weight: 194.19Product specifications:Form: White powderAssay: 98.5 - 101.0%Identity (A, B): Passes testsMelting Point: 235 - 239°CLoss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Heavy Metals: ≤ 10 ppmReadily Carbonizable Substances: Passes testOther Alkaloids: Passes testChromatographic Purity: Passes testResidual Solvents: Passes testCAS #58-08-2

12520 100 gm 1 kg

Calcium Chloride, Dihydrate

ACS Reagent GradeCaCl2•2H2OFormula Weight: 147.01Product specifications:Form: White crystalline powderAssay: 99.0 - 105.0%pH (5%, H2O, 25°C): 4.5 - 8.5Insoluble Matter: ≤ 0.01%Oxidizing Substances (NO3): ≤ 0.003%Sulfate (SO4): ≤ 0.01%Ammonium (NH4): ≤ 0.005%Barium (Ba): ≤ 0.005%Heavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 0.001%Magnesium (Mg): ≤ 0.005%Potassium (K): ≤ 0.01%Sodium (Na): ≤ 0.02%Strontium (Sr): ≤ 0.1%CAS #10035-04-8

12531 500 gm 2.5 kg

D-Calcium Pantothenate(Pantothenic Acid, Calcium Salt; Vitamin B5 )

USBioAnalyzed(C9H16NO5)2CaFormula Weight: 476.53Product specifications:Form: White powderIdentity (A, B): Passes tests[α]25

D : +25.0° to +27.5° (c=5 in H2O, dry basis)Loss on Drying: ≤ 5.0%Heavy Metals (Pb): ≤ 20 ppmOrdinary Impurities: Passes testAlkalinity: Passes testNitrogen Content (N): 5.7 to 6.0%Calcium Content (Ca): 8.2 to 8.6%Residual Solvents: Passes testCAS #137-08-6

12550 100 gm 500 gm 1 kg

We have been supplying bulk biochemicals to academic institutions, diagnostic, biotech, and pharmaceutical companies for over 40 years.

Ask us about:� Large quantities� Reserving lots and multiple lot testing� Long term supply agreements

Biochemicals


Carbenicillin, Disodium Salt

C17H16N2Na2O6SFormula Weight: 422.36Product specifications:Form: White to off-white crystalline powderAssay: 89.0 - 100.5% (dry basis)Identity: Passes testMoisture (KF): ≤ 5.0%Storage: 4°CCAS #4800-94-6

12678 1 gm 5 gm

Carboxypeptidase Y

(E.C.3.4.16.5)Source: YeastProduct specifications:Form: A useful enzyme in the sequence analysis of

proteins and peptides.Specific Activity: ≥ 100 units/mg proteinUnit Definition: One unit hydrolyzes 1 µmol of CBZ-

Phe-Leu/minute at 25°C, pH 6.5.Solubility: Soluble in water and phosphate bufferNote: Please inquire for bulk pricing.Storage: -20°C

12758 1 mg 10 mg

Casein, HammarstenULTRAPURE

Formula Weight: 75,000 - 100,000Product specifications:Form: Cream to light yellow, lyophilized powderProtein: ≥ 95%Loss on Drying: ≤ 10.0%CAS #9000-71-9

12840 100 gm 1 lb 5 lb 25 lb

Casein (High Nitrogen)

Prepared from New Zealand casein exclusivelyProduct specifications:Form: White to off-white powderProtein (dry basis): ≥ 95.0%CAS #9000-71-9

12845 5 lb 25 lb 100 lb

Casein Hydrolysate, Enzymatic DigestULTRAPURE

Product specifications:Form: PowderMoisture: ≤ 6%pH: 6.6 - 7.5Amino Nitrogen: 3.7 - 4.8%Total Nitrogen: 11.5 - 14.2%Protein (N x 6.38): 73.3 - 90.6%Ash: ≤ 6.5%CAS #65072-00-6

12855 1 lb 5 lb 25 lb

Casein, Sodium(Sodium Caseinate)

Product specifications:Form: White to off-white powderProtein (dry basis): ≥ 93%Ash: ≤ 4.2%Moisture: ≤ 5.0%pH (5%, H2O): 6.4 - 7.0CAS #9005-46-3

12865 1 lb 5 lb

Casein - Vitafree™ULTRAPURE

Product specifications:Form: Cream colored powderProtein (N x 6.25): ≥ 95.0%Loss on Drying: ≤ 12.0%Specifically prepared as a source of high quality

protein in diets needed to produce vitamin defi-ciencies.

CAS #68308-23-6

12866 5 lb 25 lb

Catalase

(E.C.1.11.1.6)Source: Bovine LiverProduct specifications:Form: Lyophilized powder, approx. one-third sucroseIdentity: By IRActivity: ≥ 3000 units/mg proteinProtein Concentration: Approx. 50%Unit Definition: One unit decomposes 1 µmol of

H2O2/minute at 25°C, pH 7.0.Storage: 4°C

12885 500 mg 1 gm 5 gm 10 gm

Cellulase

(E.C.3.2.1.4)Source: Aspergillus nigerUnit Definition: One unit produces a change in the

relative fluidity in a defined sodium carboxy-methyl cellulose substrate of 1 in 5 minutes at 40°C, pH 4.5.

Storage: 4°C

13285 100 gm

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Biochemicals


Cesium ChlorideULTRAPURE

MB GradeCsClFormula Weight: 168.36Product specifications:Form: White/clear crystalline powderAssay: 99.999%Refractive Index (50% w/w, 20°C): 1.389 - 1.391A (260 nm, 50% w/v, 1 cm): ≤ 0.02Insoluble Matter: NegligibleSilicon (Si): ≤ 1 ppmSulfate (SO4): ≤ 5 ppmBarium (Ba): ≤ 2 ppmCalcium (Ca): ≤ 3 ppmLithium (Li): ≤ 1 ppmMagnesium (Mg): ≤ 1 ppmPotassium (K): ≤ 3 ppmRubidium (Rb): ≤ 4 ppmSodium (Na): ≤ 3 ppmStrontium (Sr): ≤ 1 ppmAluminum (Al): ≤ 1 ppmChromium (Cr): ≤ 1 ppmIron (Fe): ≤ 5 ppmLead (Pb): ≤ 2 ppmManganese (Mn): ≤ 1 ppmDNase (endo): Passes testDNase (exo): Passes testRNase: Passes testApplication:� Used for density gradient high speed centrifuga-

tion separation of λ phage, M13 and plasmid DNA.

Densities in CsCl: Double-stranded DNA: density (P) = (0.098)

[G + C] + 1.660 gm/cm3 mole Sample Density RFI (Supercoiled) ds DNA 1.709 gm/cm3

RFII (Nicked) ds DNA 1.54 gm/cm3

ss DNA 1.726 gm/cm3

ss RNA 1.90 gm/cm3

Protein 1.33 gm/cm3

Note:1. Nicked DNA binds more ethidium bromide than

supercoiled DNA.2. Ethidium bromide at saturation decreases density

of ds DNA ~ 0.15 gm/cm3.3. More ethidium bromide bound = greater reduc-

tion in density.Typical Cesium Chloride Solutions: Step gradients prepared in SM* or H2O; for

(100 ml) Density CsCl SM or Refractive (P) (gm) H2O (ml) Index

1.45 60 85 1.377 1.50 67 82 1.382 1.70 75 75 1.399* SM—phage dilution buffer (100 mM NaCl, 8 mM MgSO4

•7H2O, 50 mM Tris-HCl, pH 7.5, 0.01% gelatin).

Cesium Chloride RNA Isolation Method:A. Guanidine Thiocyanate Homogenization Buffer

for RNA Isolation: 4.0 M Guanidine Thiocyanate, 0.1 M Tris, pH 7.5, 1.0% 2-mercaptoethanol

B. Cesium Chloride RNA Isolation Solution: Most researchers layer their RNA homogenates onto a cushion of 5.7 M Cesium Chloride.

C. RNA Resuspension Buffer: TE, pH 7.5.Reference:1. Schildkraut, C. L., Marmur, J. and Doty, P. (1962)

J. Mol. Biol. 4, 430-443.CAS #7647-17-8

75822 5 gm 25 gm 100 gm 500 gm 1 kg

Cetyl Trimethyl Ammonium Bromide(CTAB)

[CH3(CH2)15N(CH3)3]BrFormula Weight: 364.45Product specifications:Form: White crystalline powderAssay: ≥ 98.0% as isIdentity: By IRpH (10%, H2O): 3.0 - 7.0Moisture (KF): ≤ 1.0%Melting Point: 232 - 237°CApplications:� Cationic detergent, soluble in H2O and readily

soluble in alcohol.� Used in the preparation and purification of

genomic DNA from bacteria.� Complexes with both polysaccharide and residual

protein.� Also used for DNA mini preps for sequencing.CAS #57-09-0

13353 100 gm 500 gm 1 kg

Chicken Intestine Alkaline Phosphastase

E.C.3.1.3.1Source: Chicken IntestineProduct specifications:Form: Lyophilized powder salt freeUnit Definition: One unit hydrolyzes one µmole of

o-carboxyphenyl phosphate/minute at 25°C, pH 8.8.

Storage:-20°C, desiccated CAS #9001-78-9

10925 1 gm 5 gm

ChloramphenicolULTRAPURE

USBioAnalyzedC11H12Cl2N2O5

Formula Weight: 323.13Product specifications:Form: White to grey-white crystalline powderAssay: 97.0 - 103.0% (as is)Identity (A, B): Passes testsChromatographic Purity: Passes testMelting Point: 149 - 153°CpH (2.5%, H2O): 4.5 - 7.5Crystallinity: Passes testApplications:� Used in large-scale plasmid DNA preparation

of low to moderate copy number plasmids. Also used in high copy plasmids to limit the bulk and viscosity of the cell lysates

� Used in chloramphenicol acetyl transferase (CAT) assay

Mode of Action:Binds to the ribosomal 50S subunit and inhibits protein synthesis.Soluble in methanol, ethanol.

Stock Solution: 34 mg/ml in ethanol for plasmid DNAs

Working Concentration: 25-170 µg/ml; 100 mg/ml in ethanol for CAT assays

CAS #56-75-7

23660 25 gm 100 gm 1 kg

Cholesterol

C27H46OFormula Weight: 386.65Product specifications:Form: White to faintly yellow crystalline powderIdentity: Passes testSolubility in alcohol (1%): Passes test[α]20

D : -34° to -38° (c=2 in dioxane)Acidity: Passes testMelting Range: 147 - 150°CLoss on Drying: ≤ 0.3%Residue on Ignition: ≤ 0.1%CAS #57-88-5

13610 100 gm 1 kg

Cholesterol

Reagent GradeC27H46OFormula Weight: 386.65Storage: 4°CCAS #57-88-5

13580 5 gm 100 gm

Biochemicals


Choline Chloride

C5H14ClNOFormula Weight: 139.62Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (dry basis)Identity: By IRMoisture (KF): ≤ 0.5%Residue on Ignition: ≤ 0.05%Heavy Metals (Pb): ≤ 10 ppm1,4-Dioxane: Passes testProtect from moisture. This material is hygroscopic.CAS #67-48-1

13410 200 gm 1 kg 5 lb 25 lb

Citric Acid, Anhydrous

USBioAnalyzedC6H8O7

Formula Weight: 192.12Product specifications:Form: White crystalline powderAssay: 99.5 - 100.5% (dry basis)Clarity of Solution: Passes testColor of Solution: Passes testIdentity: By IRMoisture (KF): ≤ 1.0%Residue on Ignition: ≤ 0.1%Readily Carbonizable Substances: Passes testHeavy Metals (Pb): ≤ 0.001%Oxalate: Passes testSulfate (SO4): Passes testCAS #77-92-9

13729 500 gm 1 kg

Citric Acid, Trisodium Salt, Dihydrate(Citric Acid, Sodium Salt; Sodium Citrate, Dihydrate)

ULTRAPURE

USBioAnalyzedC6H5Na3O7

•2H2OFormula Weight: 294.10Product specifications:Form: White crystalline powderAssay: 99.0 - 100.5% (dry basis)Identity A: By IRIdentity B: Passes testLoss on Drying: 10.0 - 13.0%Tartrate: Passes testAlkalinity: Passes testHeavy Metals (Pb): ≤ 10 ppmCAS #6132-04-3

13735 1 kg 5 kg

Cocarboxylase(Thiamine Pyrophosphate Chloride)

C12H19CIN4O7P2SFormula Weight.: 460.77Product specifications:Form: White powderAssay: ≥ 93.0% (dry basis)Identity: By IRLoss on Drying (130°C): ≤ 1.5%Chloride (Cl): 7.5 - 7.9%Sulfate (SO4): ≤ 0.01%Heavy Metals (Pb): ≤ 10 ppmCAS #154-87-0

13765 25 gm 100 gm

Coenzyme A, Free Acid, Trihydrate

C21H36N7O16P3S•3H2OFormula Weight: 821.58Product specifications:Form: Lyophilized powderAssay: ≥ 75% as reduced Co-A•3H2OSpectral Ratios (pH 7.5): A250/A260: 0.78 ± 0.03 A280/A260: 0.16 ± 0.03Moisture (KF): ≤ 8%Note: The above product is available in manufactur-

ing quantities. Please inquire for bulk pricing.Storage: -20°CCAS #85-61-0

13787 50 mg 100 mg 250 mg 500 mg 1 gm

Coenzyme A, Trilithium Salt, Dihydrate

C21H33N7O16P3SLi3•2H2OFormula Weight: 821.36Product specifications:Form: White crystalline powderAssay (Enzymatic): ≥ 75%Spectral Ratios (pH 7.5): A250/A260: 0.78 ± 0.03

A280/A260: 0.16 ± 0.03Lithium (Li): 2.5 ± 1.5%Moisture (KF): ≤ 8%Note: The above product is available in manufactur-

ing quantities. Please inquire for bulk pricing.Storage: -20°C, protect from moisture.CAS #18439-24-2

13785 100 mg 250 mg 500 mg 1 gm

Coenzyme I

See β-Nicotinamide Adenine Dinucleotide, page 229

Coenzyme II

See Nicotinamide Adenine Dinucleotide Phosphate, page 230

Collagen

Source: Calf SkinProduct specifications:Form: Solution in dilute (0.075 M) acetic acid with

0.01% thimerosal as a preservative, pH 3.7.Collagen concentration: Approximately 10 mg/mlStorage: 4°C

13813 10 ml 25 ml 100 ml

Collagenase, Type I

(E.C.3.4.24.3)Source: Clostridium histolyticumDescription: Contains a mixture of collagenase,

clostripain, trypsin, caseinase, and other lytic enzymes.

Product specifications:Form: Lyophilized powderActivity:

≥ 125 units/mg powder (dry basis)≥ 0.15 Wunsch units/mg powder (dry basis)

Unit Definition: One unit liberates 1 µmol of L-Leucine equivalents from collagen in 5 hours at 37°C, pH 7.5. One unit is equal to 0.0012 Wunsch units.

Contaminating Activities:Caseinase: 200 - 1,000 units/mgClostripain: 1.0 - 3.5 units/mgTrypsin: 0.1 - 0.5 units/mg


13820 100 mg 1 gm 5 gm

Compact Reaction Columns(CRC)

With screw-on lid, Luer-lock lid, stopper and filter insertion tools.

Upper and lower filters with a pore size appropri-ate to the customer’s needs must be purchased separately.

50 columns/pk

13928 1 pk

Biochemicals


Compact Reaction Columns(CRC) without screw on lids

Stopper and filter insertion tools included.Upper and lower filters with a pore size appropriateto the customer’s needs must be purchasedseparately.100 columns/pk

13929 1 pk

CRC Lower Filters(2.7 mm)

10 μm pore size50 filters/pk

13911 1 pk



13912 1 pk



13913 1 pk

CRC Upper Filters(6.8 mm)


13914 1 pk



13918 1 pk


90 µm pore size50 filters/pk

13919 1 pk

Coomassie Stains

See Brilliant Blue G & R, page 203

Creatine Kinase

(E.C.2.7.3.2)Source: Rabbit MuscleForm: Salt-free lyophilized powderUnit Definition: One unit phosphorylates 1 µmol of

creatine/minute at 25°C, pH 9.0.Storage: -20°C, desiccatedCAS #9001-15-4

13915 100 mg 1 gm

Creatine Phosphate, Disodium Salt, Tetrahydrate(Phosphocreatine)

C4H8N3O5PNa2•4H2O

Formula Weight: 327.14Product specifications:Form: White crystalline powderAssay: ≥ 98.0%Chromatography (TLC): HomogeneousIdentity: By IRMoisture (KF): 23.5 ± 2.5%Storage: 4°CCAS #922-32-7

13920 5 gm 25 gm

CTAB

See Cetyl Trimethyl Ammonium Bromide, page 206

CTP

See Cytidine-5’-Triphosphate, page 209

Cyanocobalamin

See Vitamin B12, page 250

α-Cyclodextrin

C36H60O30

Formula Weight: 972.84Product specifications:Form: White crystalline powderAssay (HPLC): ≥ 98.0%[α]20

D : +147° to +152° (c=1 in H2O)Loss on Drying: ≤ 12%Residue on Ignition: ≤ 0.1%Chloride (Cl): ≤ 0.018%Arsenic (As): ≤ 2 ppmHeavy Metals (Pb): ≤ 10 ppmCAS #10016-20-3

13979 25 gm

L-Cysteine, Hydrochloride, Monohydrate

HSCH2CH(NH2)COOH•HCl•H2OFormula Weight: 175.63Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity: By IRChromatography: Homogeneous[α]25

D : +5.7° to +6.8° (c=8 in in 6N HCl, dry basis)Loss on Drying: 8.0 - 12.0%Residue on Ignition: ≤ 0.1%Sulfate (SO4): ≤ 0.03%Heavy Metals (Pb): ≤ 10 ppmArsenic (As): ≤ 1.5 ppmChloride (Cl): 19.8 - 20.8%Iron (Fe): ≤ 10 ppmCAS #7048-04-6

14035 25 gm 100 gm 1 kg 5 kg

L-Cystine

CP GradeC6H12N2O4S2

Formula Weight: 240.30Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity: By IR[α]20

D : -215° to -225° (c=2 in 1N HCl)Loss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 20 ppmLead (Pb): ≤ 10 ppmCAS #56-89-3

14050 50 gm 100 gm 1 kg 5 kg

Cytidine, Free Base

C9H13N3O5

Formula Weight: 243.22Product specifications:Form: White crystalline powderAssay (UV): ≥ 97.5%Identity: By IRλmax (pH 1.0): 280 nm ± 2 nmSpectral Ratios (pH 1.0): A250/A260: 0.45 ± 0.02 A280/A260: 2.10 ± 0.05Loss on Drying: ≤ 1.0%Reference constants:λmax (pH 7, NRC): 271 nmεmax (pH 7, NRC): 9.1 x 103

ε260 (pH 7, DBR): 7.55 x 103

CAS #65-46-3

14060 25 gm 100 gm

Biochemicals


Cytidine-5’-Triphosphate, Disodium Salt, Dihydrate(CTP)


Formula Weight: 527.13 (anhydrous)Formula Weight: 563.16 (dihydrate)Product specifications:Form: White powderAssay (Total UV, pH 2.0): ≥ 96.0%λmax: 280 nm ± 1 nmεmax: (pH 2.0): [13.0 ± 0.52] x 103


A280/A260: 2.10 ± 0.07Moisture (KF): ≤ 12.0%Reference constants:λmax (pH 7, NRC): 271 nmεmax (pH 7, NRC): 9.1 x 103

ε260 (pH 7, DBR): 7.4 x 103


14121 100 mg 500 mg 1 gm

Cytochrome C

Source: Horse HeartProduct specifications:Form: Reddish or dark brown crystalline powderAssay: ≥ 90.0% (dry basis)Identity: Passes testpH (1%, H2O): 5.0 - 7.0Moisture (KF): ≤ 6.0%Iron (Fe): 0.40 to 0.50%Residue on Ignition: ≤ 1.5%Solubility (10%, H2O): Clear dark red solutionStorage: -20°CCAS #9007-43-6

14102 100 mg 500 mg 1 gm

DAPI(4’,6-Diamidino-2-Phenylindole Dihydrochloride)

C16H15N5•2HCl

Formula Weight: 350.25Product specifications:Form: Yellow powderAssay (HPLC): ≥ 98%Identity: By IRSolubility (1%, H2O): ClearNitrogen: ≥ 18.0%Applications:� Cytochemical technique for demonstration of

DNA in cells infected with mycoplasmas and viruses(1)

� Interaction of DAPI with synthetic polynucleo-tides(2)

� DAPI-stained cells can be fixed for high resolution flow cytometry(3).

� Reverse fluorescent chromosome banding by double-staining with chromomycin and DAPI(4)

� Binding of DAPI to GC and mixed sequences in DNA with an intercalation of a groove-binding molecule(5)

Notes: DAPI is an excellent dye for the staining of DNA. Originally, only the specific binding to AT-base pairs without an intercalation was known(2). Later on, the intercalation into GC-base pairs was shown. The most popular application of DAPI is its use as a reagent to detect mycoplasma or virus DNA (e.g. vaccinia infection or ‘unwant-ed’ viral contamination of cell culture cells) in cell culture. Reference 1 describes a simple procedure to stain DNA with DAPI: Grow cells on a coverslip in a cell culture dish. Wash the coverslip once with PBS. Incubate the cells on the coverslip at 37°C for 15 - 30 minutes in 0.1 µg DAPI/ml PBS. Wash the coverslip in PBS and put it up-side-down on a slide. Examine the cells under a micro-scope (excitation: 365 nm; emission maximum at 450 nm). Prolonged incubation with DAPI increases the nuclear fluorescence; shorter incu-bation time leads to a weaker nuclear staining, which facilitates the examination of the cytoplas-mic fluorescence.

DAPI can be dissolved in water, PBS, 5 mM Tris-HCl (pH 7.6)/8 mM NaCl, citrate or phosphate buf-fer(3). The solubility is approximately 2.5%. Stock solutions are prepared at concentrations ranging from 1-2 mg/ml and the working dilution is 1:1000. DAPI solution is stable at room tempera-ture for 1 - 2 weeks(3), at 4°C up to 6 months, and at -20°C for 6 - 12 months. DAPI has hydrolyzed if the solution becomes turbid. DAPI bleaches quickly in contact with light, even though it is quite stable against UV-light.

References:1. Russel, W. C., Newman, C. and Williamson, D. H.

(1975) Nature 253, 461-462.2. Kapuscinski, J. and Szer, W. (1979) Nucleic Acids

Res. 6, 3519-3534. 3. Otto, F. (1990) Methods Cell Biol. 33, 105-110.4. Schweizer, D. (1976) Chromosoma. 29, 307-324.5. Wilson, W. D., Tanious, F. A., Barton, H. J.,

Jones, R. L., Fox, K., Wydra, R. L. and Strekowski, L. (1990) Biochem. 11, 8452-8461.


14564 10 mg 50 mg

Denhardt’s Solution, 50XULTRAPURE

MB GradeForm: 50X solution of 1% acetylated bovine serum

albumin (0.2 µm filtered), 1% Ficoll (autoclaved before use), and 1% PVP-90 (autoclaved before use). Dispensed into autoclaved bottles.

Contaminant Testing:DNase (endo): Passes testDNase (exo): Passes testRNase: Passes testProtease: Passes test

Storage: -20°CApplication: � For use as a blocking reagent in nucleic acid

hybridization techniquesFormamide Hybridization Buffer: 50% Formamide,

5X SSC, 0.5% SDS, 5X Denhardt’s Solution, 100 µg/ml sonicated fish sperm DNA

Reagent Amount for every 100 mlFormamide* 50 ml20X SSC* 25 ml20% SDS* 2.5 ml50X Denhardt’s Solution* 10 ml10 mg/ml fish DNA 1.0 mlWater* up to 100 ml*Denotes Ultrapure Reagent available.Reference:1. Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989)

Molecular Cloning, 2nd Edition, Cold Spring Harbor Laboratory.

70468 50 ml

2’-Deoxyadenosine

C10H13N5O3•H2O

Formula Weight: 269.26Product specifications:Form: White to off-white crystalline powderAssay (Total UV): ≥ 98%Identity: By IRλmax: 258-260 nmSpectral Ratios: A250/A260: 0.77 ± 0.04 A280/A260: 0.15 ± 0.05Moisture (KF): ≤ 8.0%Reference constants:λmax (pH 7, NRC): 260 nmεmax (pH 7, NRC): 14.9 x 103

ε260 (pH 7, DBR): 15.2 x 103

CAS #16373-93-6

14235 5 gm 25 gm

Biochemicals


2’-Deoxyadenosine-5’-Triphosphate, Sodium Salt, 100 mM Solution(dATP)

ULTRAPURE

C10H12N5O12P3Na4

Formula Weight: 579.11Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λ max: 259 nm ± 1 nmSpectral Ratios: A250/A260: 0.80 ± 0.03 A280/A260: 0.12 ± 0.02Functional Tests: Functionally tested in long PCR to



Deoxycholic Acid, Sodium SaltSodium Deoxycholate

C24H39NaO4

Formula Weight: 414.55Product specifications:Form: White to off-white powderAssay (HPLC): ≥ 98% (dry basis)Identity: By IRLoss on Drying (4 hours, 140°C, vacuum): ≤ 6%Sodium Cholate: ≤ 2%CAS #302-95-4

21026 25 gm 100 gm

2’-Deoxycytidine, Hydrochloride, Synthetic

C9H13N3O4•HCI

Formula Weight: 263.68Product specifications:Form: White crytalline powderAssay (Total UV): ≥ 98% (dry basis)Identity: By IRλmax: 280 ± 2 nmεmax (pH 1.0): 13.2 x 103 ± 2%Spectral Ratios (pH 1.0): A250/A260: 0.43 ± 0.03 A280/A260: 2.10 ± 0.05Moisture (KF): ≤ 1%Heavy Metals: ≤ 10 ppmCAS #3992-42-5

14284 1 gm 5 gm 10 gm 100 gm

2’-Deoxycytidine-5’-Triphosphate, Sodium Salt, 100 mM Solution(dCTP)

ULTRAPURE

C9H12N3O13P3Na4

Formula Weight: 555.08Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λ max (pH 2.0): 280 nm ± 1 nmSpectral Ratios (pH 2.0): A250/A260: 0.45 ± 0.03 A280/A260: 2.10 ± 0.15 A290/A260: 1.60 ± 0.10Functional Tests: Functionally tested in long PCR to



2’-Deoxyguanosine

C10H13N5O4•H2O

Formula Weight: 285.26Product specifications:Form: White granular powderAssay (HPLC): ≥ 97%Identity: By IRChromatography (TLC): Homogeneousλmax: 253 ± 2 nmεmax: 12800-14300Spectral Ratios (pH 7.0): A250/A260: 1.09 - 1.21

A280/A260: 0.63 - 0.71Moisture (KF): ≤ 9%Reference constants:λmax (pH 7, NRC): 254 nmεmax (pH 7, NRC): 13.0 x 103

CAS #961-07-9

14300 5 gm 25 gm 100 gm

2’-Deoxyguanosine, Monohydrate Synthetic

C10H13N5O4•H2O

Formula Weight: 285.26Product specifications:Form: White crystalline powderAssay: ≥ 98% (dry basis)Chromatography (TLC): HomogeneousIdentity: By IRλmax (pH 7.0): 253 ± 2 nmεmax (pH 7.0): (13.7 x 103) ± 4%Spectral Ratios (pH 7): A250/A260: 1.12 - 1.18 A280/A260: 0.65 - 0.69Moisture (KF): ≤ 7.0%CAS #961-07-9

14304 1 gm 5 gm 10 gm 100 gm 1 kg

7-deaza-2’-Deoxyguanosine-5’-Triphosphate, Lithium Salt, 10 mM Solution(7-deaza-dGTP)

C11H15N4O13P3Li2Formula Weight: 518.10Product specifications:Form: 10 mM aqueous solutionAssay (HPLC): ≥ 95%pH: 7.0λmax (pH 7.5): 259 nm ± 1 nmFunctional Tests: Meets or exceeds criteria for high


70065 2 µmol

2’-Deoxyguanosine-5’-Triphosphate, Sodium Salt, 100 mM Solution(dGTP)

ULTRAPURE

C10H12N5O13P3Na4




Biochemicals


2’-Deoxynucleoside-5’-Triphosphates, Sodium Salt, 100 mM Solutions, Sets of Four(dNTPs)

ULTRAPURE

Form: 100 mM aqueous solutions, pH 7.5; dATP, dCTP, dGTP, dTTP

Functional Tests: Functionally tested in long PCR to generate a 20.7 kb fragment.

Storage: -20°C

4 x 25 µmol (250 µl) each dNTP per pack77100 1 pk4 x 100 µmol (1 ml) each dNTP per pack77328 1 pk4 x 500 µmol (5 ml) each dNTP per pack77128 1 pk

2’-Deoxynucleoside-5’-Triphosphate Mixes

See PCR Nucleotide Mixes, page 232

2-Deoxy-D-RiboseULTRAPURE

C5H10O4

Formula Weight: 134.13Product specifications:Form: White powderAssay: ≥ 98.0%[α]20

D : -57.3° ± 1.5°(c=1 in H2O)Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.5%Heavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 5 ppmCAS #533-67-5

14420 5 gm 25 gm 100 gm

2’-Deoxythymidine-5’-Triphosphate, Sodium Salt, 100 mM Solution(dTTP)

ULTRAPURE

C10H13N2O14P3Na4




2’-Deoxyuridine-5’-Triphosphate, Sodium Salt, 100 mM Solution(dUTP)

ULTRAPURE

C9H11N5O12P3Na4

Formula Weight: 556.1Product specifications:Form: 100 mM aqueous solutionAssay (HPLC): ≥ 99%pH: 7.5 ± 0.2λmax (pH 7.5): 262 nm ± 1 nmAbsorbance ratios: A250/A260: 0.74 ± 0.03 A280/A260: 0.35 ± 0.03Functional Test: Functionally tested in PCRStorage: -20°C


DEPC

See Diethyl Pyrocarbonate, page 212

DextranULTRAPURE

M.W.: 60,000 - 90,000Product specifications:Form: White powderpH (6% w/v in KCl): 4.5 - 7.0Loss on Drying: ≤ 7.0%Residue on Ignition: ≤ 1.9%CAS #9004-54-0

14495 100 gm 1 kg

Dextran Sulfate, Sodium Salt

M.W.: 40,000 - 50,000Product specifications:Form: Off white to light yellow powderLoss on Drying: ≤ 10.0%pH (1%, H2O): 5.0 - 7.5Specific Viscosity (25°C in 1.0 M NaCl): 0.05 - 0.13Sulfur: 17.0 - 20.0%Glucose: 35.0 - 48.0%Residue on Ignition: 35.0 - 50.0%CAS #9011-18-1

14489 10 gm 50 gm 100 gm

Dextran Sulfate, Sodium SaltULTRAPURE

MW: 500,000*Product specifications:Form: White to off-white powderLoss on Drying: ≤ 10%pH (1%, H2O): 5.0-7.5Residue on Ignition: 40 - 50%* Note: Approximate molecular weight of dextran used in the production of the sulfate. Average MW of dextran sulfate sodium salt is approx. 1,400,000.

CAS #9011-18-1

70796 10 gm 50 gm

D-(+)-Dextrose, Anhydrous(α-D-(+)-Glucose)

CH2OH(CHOH)4CHOFormula Weight: 180.16Product specifications:Form: White crystalline powderIdentity: By IR[α]25

D : +52.6° to +53.2° (dry basis, c=10 in 0.012N NH4OH)

Moisture (KF): ≤ 1.0%CAS #50-99-7

14535 5 lb 25 lb 100 lb

Biochemicals


D-(+)-Dextrose, Monohydrate(α-D-(+)-Glucose)

C6H12O6•H2O

Formula Weight: 198.17Product specifications:Form: White crystalline powderIdentity: By IR[α]25

D : +52.6° to +53.2° (dry basis, c=10 in 0.12N NH4OH)

Moisture (KF): ≤ 9.5%Storage: Ambient, in tightly closed containers.CAS #50-99-7

14536 10 kg

Diaphorase

(E.C.1.8.1.4)Source: Clostridium kluyveriForm: Lyophilized powderUnit Definition:

One DCPIP unit is that amount of enzyme which causes a decrease in absorbance of 1.0 per min-ute at 25°C, pH 7.5, with 2,6-dichlorophenolin-odophenol as the chromogen.

One MTT unit is that amount of enzyme which reduces 1 µmol of MTT per minute at 30°C under stated assay conditions.

Activity (DCPIP units/mg): ≥ 30 DCPIP units/mg dry weight

Activity (MTT units/mg): ≥ 3.9 MTT units/mg dry weight


14615 600 units 1,800 units

2’,3’-Dideoxyadenosine-5’-Triphosphate(ddATP)

ULTRAPURE

C10H12N5O11P3Na4

Formula Weight: 563.11Product specifications:Assay (FPLC): ≥ 99%pH: 7.5 ± 0.2λmax: 259 nm ± 1 nmSpectral Ratios (pH 7.5): A250/A260: 0.80 ± 0.03 A280/A260: 0.12 ± 0.02Functional Test: Meets or exceeds criteria for high



2’,3’-Dideoxycytidine-5’-Triphosphate(ddCTP)

ULTRAPURE

C9H12N3O12P3Na4

Formula Weight: 539.08Product specifications:Assay (FPLC): ≥ 99%pH: 7.5 ± 0.2λmax (pH 2.0): 280 nm ± 1 nmSpectral Ratios (pH 2.0): A250/A260: 0.45 ± 0.03 A280/A260: 2.20 ± 0.20 A290/A260: 1.70 ± 0.20Functional Test: Meets or exceeds criteria for high



2’,3’-Dideoxynucleoside-5’-Triphosphates, 10 mM Solutions, Sets of Four(ddNTPs)

ULTRAPURE

Form: 10 mM aqueous solutions, pH 7.5; ddATP, ddCTP, ddGTP, ddTTP

Storage: -20°C

4 x 0.5 µmol (50 µl) each ddNTP per pack77126 1 pk4 x 1.0 µmol (100 µl) each ddNTP per pack77125 1 pk

2’,3’-Dideoxynucleoside-5’-Triphosphates, 100 mM Solutions, Set of Four(ddNTPs)

ULTRAPURE

Form: 100 mM aqueous solutions, pH 7.5; ddATP, ddCTP, ddGTP, ddTTP

Storage: -20°C

4 x 4 µmol (40 µl) each ddNTP per pack77335 1 pk

2’,3’-Dideoxythymidine-5’-Triphosphate(ddTTP)

ULTRAPURE

C10H13N2O13P3Na4

Formula Weight: 554.10Product specifications:Assay (FPLC): ≥ 99%pH: 7.5 ± 0.2λmax (pH 7.5): 267 nm ± 1 nmSpectral Ratios (pH 7.5): A250/A260: 0.65 ± 0.03 A280/A260: 0.83 ± 0.03Functional Tests: Meets or exceeds criteria for high



Diethyl Pyrocarbonate(DEPC)

ULTRAPURE

O(COOC2H5)2

Formula Weight: 162.14Product specifications:Form: Clear, colorless liquidAssay: ≥ 97%Density (20°C): 1.121 ± 0.005Refractive Index (20°C): 1.398 ± 0.005Applications:� A strong, nonspecific inhibitor of ribonuclease� Reacts with the N1 of histidine residues in pro-

teins under mild conditions.� Used to cross-link and modify proteins by eth-

oxyformylation of NH2 and imidazole groups in proteins.

� Reacts with amines and thus cannot be used with Tris Buffer.

References:1. Tsurushiin, S., Hiramatsu, A., Inamasu, M. and

Yasunobu, K. T. (1975) Biochim. Biophys. Acta. 400, 451-460.

2. Meyer, S. E. and Cromartie, T. H. (1980) Biochemistry 19, 1874-1881.

3. Berger, S. L. (1975) Anal. Biochem. 67, 428-437.4. Leonard, N. J., Mcdonald, J. J., Henderson, R. E.

and Reichmann, M. E. (1971) Biochemistry 10, 3335-3342.

5. Roberts, M. F., Deems, R. A., Mincey, T. C. and Dennis, E. A. (1977) J. Biol. Chem. 252, 2405-2411.

Storage: -20°CCAS #1609-47-8

14710 25 ml 100 ml

Biochemicals


Dihydrostreptomycin Sulfate

USBioAnalyzed(C21H41N7O12)2

•3H2SO4

Formula Weight: 1461.42Product specifications:Form: White to off-white powderActivity: ≥ 650 µg of C21H41N7O12/mg (as is)Identity (A, B): Passes testspH (200 mg dihydrostreptomycin/ml): 4.5 - 7.0Loss on Drying: ≤ 5.0%Streptomycin: ≤ 3.0%CAS #5490-27-7

14765 100 gm

Dimedone(5,5-Dimethyl-1,3-Cyclohexanedione)

C8H12O2

Formula Weight: 140.18Product specifications:Form: White to off-white crystalline powderAssay: ≥ 99%Identity: By IRMelting Point: 145 - 151°CSolubility (5% EtOH): Passes testStorage: Ambient, in tightly closed containers.CAS #126-81-8

14850 100 gm 1 kg

p-Dimethylamino Benzaldehyde

(CH3)2NC6H4CHOFormula Weight: 149.19Product specifications:Form: White to off-white powderIdentity: By IRChromatography (TLC): HomogeneousMelting Point: 70 - 75°CCAS #100-10-7

14868 100 gm 500 gm

p-Dimethylamino Cinnamaldehyde

(CH3)2NC6H4CH:CHCHOFormula Weight: 175.23Product specifications:Form: Bright yellow/gold crystalline powderAssay: ≥ 98.0%Identity: By IRMelting Point: 137 - 142°CCAS #6203-18-5

14872 25 gm

Dithioerythritol(DTE)

ULTRAPURE

C4H10O2S2

Formula Weight: 154.25Form: White crystalline powderChromatography (TLC): HomogeneousAssay (I2): ≥ 99%Melting Point: ≥ 81°CA (283 nm, 0.02 M, H2O): ≤ 0.04Storage: -20°C, protect from moisture.CAS #6892-68-8

15385 5 gm 25 gm 100 gm

Dithiothreitol(DTT; Cleland’s Reagent)

ULTRAPURE

MB GradeC4H10O2S2

Formula Weight.: 154.25Product specifications:Form: White crystalline powderAssay (SH): ≥ 99.5%A (280 nm, 0.1 M, 1 cm): ≤ 0.08A (260 nm, 0.1 M, 1 cm): ≤ 0.40Melting Point: 40 - 43°COxidized DTT: ≤ 0.3%DNase: Passes testRNase: Passes testProtease: Passes testApplication:� Used as a protective agent for SH groups.Reference:1. Cleland,W. W. (1964) Biochemistry 3, 480-482.Storage: -20°CCAS #3483-12-3

15397 1 gm 5 gm 25 gm 100 gm

Dithiothreitol(DTT; Cleland’s Reagent)

C4H10O2S2

Formula Weight: 154.25Product specifications:Form: White crystalline powderAssay: ≥ 97%Identity: By IRMelting Point: ≥ 38°COxidized DTT: ≤ 1%Storage: -20°C, in tightly closed containers.CAS #3483-12-3

15395 5 gm 25 gm 100 gm

Dithiothreitol (DTT), 0.1 M SolutionULTRAPURE

MB GradeProduct specifications:Form: 0.1 M solution of DTT in high purity dH2O.

Solution is 0.2 µm filtered and dispensed into vials.

Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testStorage: -20°C

70726 150 µl 1 ml 5 ml

DNA, Calf ThymusDNA, E. coliDNA, Fish SpermDNA, Fish Sperm, Activated DNA-Cellulose, Double-Stranded(dsDNA-Cellulose)DNA-Cellulose, Single-Stranded(ssDNA-Cellulose)

See Molecular Biology Products & Kits section, pages 128–129

DNase I

(E.C.3.1.21.1)Source: Bovine pancreasForm: Lyophilized powderActivity:

>90,000 Dornase units/mg>2,500 Kunitz units/mg

Unit Definition: One Kunitz unit causes an increase in absorbance of 0.001 per minutes per ml at 260 nm at 25°C, pH 5, using the assay of Kunitz(1). One Kunitz unit = 35 Dornase units.

Reference:1. Kunitz (1950) J. Gen. Physiol. 33, 349-363.Storage: 4°C

14340 10 mg 20 mg

Dodecyl Sulfate, Lithium Salt

See Lithium Dodecyl Sulfate, page 225

Dodecyl Sulfate, Sodium Salt

See Sodium Dodecyl Sulfate, page 240

Biochemicals


N-Dodecyl Trimethylammonium Bromide(DTAB)

C15H34BrNFormula Weight: 308.34Product specifications:Form: White powderMelting Point: 239 - 247°CCAS #1119-94-4

18218 10 gm 50 gm 250 gm

DPN

See Nicotinamide Adenine Dinucleotide, page 229

DTAB

See N-Dodecyl Trimethylammonium Bromide, page 214

DTE

See Dithioerythritol, page 213

DTT

See Dithiothreitol, page 213

EDTA

See Ethylenediamine Tetraacetic Acid, page 215

Egg White

See Albumin-Egg, page 199

EGTA

See Ethylene Glycol-o-o’-bis (2-aminoethyl)-N,N,N’,N’,-Tetraacetic Acid, page 215

Elastase

(E.C.3.4.21.11)Source: Porcine PancreasForm: Lyophilized, salt free powderUnit Definition: One unit solubilizes 1 mg of elas-

tin/20 minutes at 37°C, pH 8.8.Storage: 4°CCAS #9004-06-2

15475 5 mg 10 mg 20 mg 100 mg

Enolase

(E.C.4.2.1.11)Source: YeastProduct specifications:Form: 50% glycerol solutionSpecific Activity: ≥ 40 units/mg proteinContaminating Activities:

Phosphoglyceromutase: ≤ 0.02%Pyruvate Kinase: ≤ 0.02%

Glycerol: 50%K-PO4: 0.1 M ± 0.05 MpH: 7.0Unit Definition: One unit oxidizes 1 µmol of NADH/

minute at 25°C, pH 7.5.Storage: -20°C

15515 10,000 units

Ergocalciferol

USBioAnalyzedC28H44OFormula Weight: 396.65Product specifications:Form: White to light yellow powderIdentification (A, B, C, D,): Passes testAssay: 97.0 - 103.0% (as is)[α]25

D : +103° to +106° (c= 1.5 in EtOH)Melting Range: 115 - 119°CReducing Substances: Passes testStorage: 4°C, protected from light sealed under

nitrogen.

15562 5 gm 25 gm 100 gm

Esculin

C15H16O9

Formula Weight: 340.28 (anhydrous)Product specifications:Form: White to off-white powderAssay: 97.5 - 102.5%Identity: By IR[α]20

D : -84° to -87° (c=2 in Dioxane:H2O/1:1, dry basis)

Moisture (KF): 6.5 - 8.0%CAS #66778-17-4

15590 100 gm

Ethidium BromideULTRAPURE

C21H20BrN3

Formula Weight.: 394.31Product specifications:Form: Purple/maroon crystalline powderAssay: ≥ 98.0%Residue on Ignition: ≤ 0.2%Ammonium Bromide: ≤ 0.1%Moisture (KF): ≤ 10.0%Applications:� A fluorescent dye that binds to DNA by intercalat-

ing between the bases, causing the double helix to unwind.

� Used to detect both single-and double-stranded DNA and RNA.

� Used in DNA isolation procedures and in gel elec-trophoresis.

Stock Solution: 10 mg/ml 100 mg Ethidium Bromide 10 ml H2O Filter and store solution protected from light.Working Concentration: 0.5 µg/ml for gel staining.Note: The electrophoretic mobility of linear double-

stranded DNA is reduced by 15% in the presence of ethidium bromide.

CAS #1239-45-8

32813 1 gm 5 gm 25 gm

Ethidium Bromide DropsULTRAPURE

Form: Ethidium bromide is supplied in a convenient, ready-to-dispense dropper format for gel electro-phoresis applications. One drop contains 40 µl or 25 µg of ethidium bromide.

Concentration: 0.625 mg/ml in water.Testing: Functionally tested for use in gel electro-

phoresis.To Use: Simply add one drop for every 50 ml of aga-

rose and/or buffer for a working concentration of 0.5 µg/ml.

75816 5 ml

Biochemicals


Ethylenediaminetetraacetic Acid, (EDTA), 0.5 M SolutionULTRAPURE

MB GradeForm: 0.5 M solution of EDTA•Na2 in high purity

dH2O, pH 8.0. Solution is 0.2 µm filtered and dis-pensed into durable, square Nalgene bottles.

pH: 8.0 ± 0.5Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testApplication: � Chelating agent used in many enzyme buffers

and, at higher concentrations, as an enzyme inac-tivator

15694 100 ml 500 ml

Ethylenediaminetetraacetic Acid, (EDTA) Disodium Salt, DihydrateULTRAPURE

C10H14N2Na2O8•2H2O

Formula Weight: 372.24Product specifications:Form: White crystalline powderAssay: 99.0 - 101.0%A (260 nm, 0.1 M, 1 cm): ≤ 0.2A (280 nm, 0.1 M, 1 cm): ≤ 0.05Insoluble Matter: ≤ 0.005%pH (5%, H2O): 4.0 - 6.0Nitrotriacetic Acid: ≤ 0.1%Heavy Metals (Pb): < 0.005%Iron (Fe): ≤ 0.01%Arsenic (As): ≤ 1 ppmCalcium (Ca): Passes testSolubility (0.1 M, H2O): Completely clear and color-

lessContaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testApplication: � Chelating agent used in many enzyme buffers

and as an enzyme inactivator at high concentra-tions

Stock Solution (0.5 M): 186.1 gm EDTA H2O to 800 ml Titrate with approximately 20 gm of NaOH pellets

to pH 8.0. Bring solution to 1 liter with water and sterilize by autoclaving.

CAS #6381-92-6

15701 100 gm 500 gm 1 kg

Ethylenediaminetetraacetic Acid, (EDTA) Disodium Salt, Dihydrate

ACS Reagent GradeC10H14N2O8Na2O8

•2H2OFormula Weight: 372.24Product specifications:Form: White crystalline powderAssay: 99.0 - 101.0%pH (5%, H2O, 25°C): 4.0 - 6.0Insoluble Matter: ≤ 0.005%Nitrilotriacetic Acid: ≤ 0.1%Heavy Metals (Pb): ≤ 0.005%Iron (Fe): ≤ 0.01%A (260 nm, 0.1 M, 1 cm): ≤ 0.2A (280 nm, 0.1 M, 1 cm): ≤ 0.05Solubility: (0.1 M, H2O): Clear and colorlessCAS #6381-92-6

15699 500 gm 1 kg 5 kg

Ethylenediaminetetraacetic Acid, (EDTA) Disodium Salt, Dihydrate

C10H14N2O8Na2•2H2O

Formula Weight: 372.24Product specifications:Form: White crystalline powderAssay: 99.0 - 101.0% (as disodium dihydrate)Identity: By IRSolubility (5%, H2O): Clear, colorless to slightly yel-

lowpH (5%, H2O): 4.0 - 6.0pH (1%, H2O): 4.0 - 6.0Note: This product is a general quality laboratory

chelating reagent suitable for most educational purposes. 0.1 M stock solutions of this grade may be pale yellow in color. For demanding research applications and for use with sensitive reagents, we recommend Ultrapure Grade (PN 15701) or ACS Reagent Grade EDTA (PN 15699).

CAS #6381-92-6

15697 1 kg

Ethylene Diamine Tetraacetic Acid, (EDTA) Tetrasodium Salt, DihydrateULTRAPURE

C10H12N2O8Na4•2H2O

Formula Weight: 416.20 (dihydrate)Product specifications:Form: White crystalline powderAssay: 99.0 - 101.0% (as is)Identity: By IRpH (1%, H2O): 10.7 - 11.7pH (5%, H2O): 10.7 - 12.0CAS #10378-23-1

15700 1 kg

Ethylene Glycol-O,O’-Bis (2 Amino-ethyl)-N,N,N’,N’,-Tetraacetic Acid (EGTA)ULTRAPURE

C14H24N2O10

Formula Weight: 380.35Product specifications:Form: White powderAssay: ≥ 99%Spectral Ratios (280 nm, 0.1 M in 1N NaOH, 1 cm):

≤ 0.10Identity: By IRLoss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Chloride (Cl): ≤ 0.005%Sulfate (SO4): ≤ 0.02%Cadmium (Cd): ≤ 5 ppmCobalt (Co): ≤ 5 ppmCopper (Cu): ≤ 5 ppmIron (Fe): ≤ 5 ppmLead (Pb): ≤ 5 ppmNickel (Ni): ≤ 5 ppmZinc (Zn): ≤ 5 ppmApplication: � Chelating agent useful in the determination of

calcium in the presence of magnesiumReference:1. Schmid, R. W. and Reilley, C. N. (1957) Anal.

Chem. 29, 264-268.CAS #67-42-5

15703 10 gm 50 gm 100 gm

Fetal Bovine Serum (FBS)

Cell Culture GradeProduct specifications:Form: Clear straw to amber liquidTotal Protein: 3.0 - 4.0 gm/dlHemoglobin: ≤ 20 mg/dlpH: 6.7 - 8.0Endotoxin: ≤ 10 EU/mlSterility: SterileMycoplasma: None detectedVirally tested and found negative for BVD, IBR, and

PI3. USB Fetal Bovine Serum is an excellent supplement

to culture media.Storage: -20°C

15762 500 mI

Biochemicals


Flavin Adenine Dinucleotide, Disodium Salt, Hydrate(FAD)

C27H31N9O15P2Na2

Formula Weight:829.51 (anhydrous)883.56 (trihydrate)

Product specifications:Form: Orange-yellow crystalline powderAssay (UV, 450 nm, pH 7.0): ≥ 95.0% (dry basis,

disodium salt)Moisture (KF): ≤ 8.0%Note: This product contains a varying amount of

moisture ranging from a dihydrate to a tetrahy-drate at the time of packaging.

Storage: 4°C, protect from light, dessicate.CAS #84366-81-4

15785 250 mg 500 mg 1 gm

Folic Acid(Pteroylglutamic Acid)

USBioAnalyzedC19H19N7O6

Formula Weight: 441.40Product specifications:Form: Yellow to yellow-orange crystalline powderAssay: 97.0 - 102.0% (dry basis)Chromatographic Purity: Passes testIdentity: Passes testMoisture (KF): ≤ 8.5%Residue on Ignition: ≤ 0.3%Organic Volatile Impurities: Meets requirementsStorage: Ambient, protect from light.CAS #59-30-3

15880 25 gm 100 gm 1 kg

FormamideULTRAPURE

Low ConductivityHCONH2

Formula Weight: 45.04Form: Clear, colorless liquidProduct specifications:Assay: ≥ 99%Refractive Index (20°C): 1.447 ± 0.001Density (20°C): 1.133 ± 0.001A (280 nm, 100%, 1 cm): ≤ 0.15A (260 nm, 100%, 1 cm): ≤ 2.3pH (50% v/v with H2O): 6.0 - 8.0Freezing Point: 2.0 - 3.0°CConductivity: ≤ 100 µmhos/cmCopper (Cu): ≤ 0.2 ppmIron (Fe): ≤ 1 ppmLead (Pb): ≤ 1 ppmZinc (Zn): ≤ 1 ppmStorage: 4°CApplications:� For use in nucleic acid hybridization buffers, spe-

cialized sequencing gels, and a variety of other molecular biology applications to reduce the Tm of nucleic acid hybrids

� Used to achieve stronger denaturing conditions in a sequencing gel along with 7-deaza-dGTP to resolve sequencing gel compressions.

To prepare a formamide sequencing gel: Place 40 ml Formamide in a beaker with mag-

netic stir bar. Acrylamide 7.6 gm N-N’-Methylene-bis-Acrylamide 0.4 gm Urea 42.0 gm 10X TBE Buffer 10 ml1. Cover with parafilm and warm in a 65°C water

bath with stirring until dissolved. This takes 30- 60 minutes and the mixture reaches a tempera-ture of about 50°C.

2. When completely dissolved, add water to a total volume of 100 ml.

3. Cover and continue stirring at room temperature until it reaches < 30°C.

4. Vacuum filter with a nitrocellulose filter.5. Add 1 ml of 10% ammonium persulfate and

0.15 ml of TEMED. Pour immediately. (Pouring this viscous mixture requires nearly a vertical angle for the glass plates.) The gel should be polymerized within 30 minutes.

Formamide gels should be run at the same power (wattage) as regular gels. This requires a higher voltage (60% higher) than normal gels, and DNA migrates about half as fast.

Soaking the gel in 5% acetic acid, 20% methanol helps prevent swelling. Soak 15 minutes for a 0.4 mm thick gel and dry normally.

Compression artifacts are caused whenever stable secondary structures exist in the DNA under the conditions prevailing in the gel matrix during electrophoresis.

The folded structure runs faster through the gel matrix than equivalent unfolded DNA, catch-ing up with the smaller DNA in the sequence. Currently, gel compression artifacts are elimi-nated in one of two ways. One is to change to a stronger denaturing condition, adding formamide as shown here, and the other is to use an analog of dGTP (dITP or 7-deaza-dGTP) during synthesis.

Formamide Hybridization Buffer: 50% Formamide, 5X SSC, 0.5% SDS, 5X

Denhardt’s Solution, 100 μg/ml sonicated fish sperm DNA

Reagent Amount for every 100 ml Formamide* 50 ml 20X SSC* 25 ml

20% SDS* 2.5 ml50X Denhardt’s Solution* 10 ml

10 mg/ml fish DNA 1.0 ml Water* up to 100 ml*Denotes Ultrapure Reagent available.References:1. Sarkar, G., Kapelner, S. and Sommer, S. S. (1990)

Nucl. Acids Res. 18, 7465.2. Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989)

Molecular Cloning, 2nd Edition, Cold Spring Harbor Laboratory.

CAS #75-12-7

75828 500 gm 1 kg

β-D-Fructose(β-D-Levulose)

C6H12O6

Formula Weight: 180.16Product specifications:Form: White crystalline powderAssay: 100 ± 2% (dry basis)Identity: By IRLoss on Drying: ≤ 0.5%Glucose: ≤ 0.5%Hydroxymethylfurfural: ≤ 0.1%Residue on Ignition: ≤ 0.5%Chloride (Cl): ≤ 0.018%Sulfate (SO4): ≤ 0.025%Arsenic (As): ≤ 1 ppmHeavy Metals (Pb): ≤ 5 ppmCAS #57-48-7

18425 1 kg

D-Fructose-6-Phosphate, Disodium Salt

C6H11Na2O9PFormula Weight: 304.10 (anhydrous) 322.11 (monohydrate)Practically free of Fructose-1,6-DiphosphateStorage: -20°CCAS #26177-86-6

15929 1 gm 5 gm

Biochemicals


G-418 Sulfate(Geneticin®, Antibiotic G-418)

ULTRAPURE

C20H40N4O10•2H2SO4

Formula Weight: 692.71Product specifications:Form: White powderPotency: ≥ 730 units/mg (dry basis)[α]25

D : +104° to 121° (c=1 in H2O)Moisture (KF): ≤ 6%Storage: 4°C

11379 500 mg 1 gm 5 gm

D-(+)-Galactose

C6H12O6

Formula Weight: 180.16Product specifications:Form: White to off-white powderAssay (HPLC): ≥ 99%Identity: By IR[α]20

D : ≥ +77° (c=4 in H2O and NH3)Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.5%CAS #59-23-4

15995 500 gm 1 kg 5 kg

GDP

See Guanosine-5’-Diphosphate, page 221

Gel Electrophoresis Reagents

Ethidium Bromide Drops, page 214Glycerol Tolerant Gel (GTG) Buffer, 20X Solution,

page 219RapidRun Agarose Buffer, 20X Solution, page 238TAE Buffer, 10X Solution, page 243TAE Buffer, 50X Solution, page 243TBE Buffer, 5X Solution, page 243TBE Buffer, 10X Ready-Mixed Powder, page 243Tris-Glycine (TG) Buffer, 10X Solution, page 247Tris-Glycine-SDS (TGS) Buffer, 10X Solution,

page 247

Gel Mixes and Solutions


RapidGel 40% Liquid Acrylamide Stock Solution, page 237

RapidGel-XL 6% DNA Sequencing Gel Solution, page 238

Stop Solution, page 242

Geneticin Disulfate

See G-418 Sulfate, page 217

Gentamicin Sulfate

USBioAnalyzedProduct specifications:Form: White to buff-colored powderActivity:

≥ 590 µg of gentamicin/mg (dry basis)≥ 484 µg of gentamicin/mg (as is)

Identity (A, B): Passes testspH (4%, H2O): 3.5 - 5.5[α]25

D : +107° to +121° (c=1 in H2O)Loss on Drying: ≤ 18.0%Residue on Ignition: ≤ 1.0%Gentamycin’s content: C1: 25 - 50% C2 + C2a: 25 - 55% C1a: 10 - 35%Limit of Methanol: ≤ 1.0%Mode of Action: Bactericidal, inhibits normal protein

synthesis.Applications:� Very stable� Autoclavable� Requires no refrigeration� Effective when combined with an antibiotic that

interferes with cell wall synthesis.CAS #1405-41-0

16051 1 gm 5 gm 25 gm

γ-Globulins, Fraction II

Source: Bovine BloodProduct specifications:Form: Lyophilized powderAssay: ≥ 98%Identity: By immuno-electrophoresisStorage: 4°CCAS #68476-36-8

16021 10 gm 25 gm

α-D-(+)-Glucose

See Dextrose, page 211

Glucose Oxidase

(E.C.1.1.3.4)Source: Aspergillus nigerForm: Stabilized, aqueous solutionActivity: Approx. 260 units/mg proteinProtein Concentration: Approx. 20 mg protein/mlContaminating Activities:

Amylase: ≤ 0.05%Saccharase: ≤ 0.05%Maltase: ≤ 0.05%

Unit Definition: One unit oxidizes 1 µmol of glucose/minute at 25°C, pH 7.0.

Storage: 4°C

16146 50,000 units 250,000 units 1 mu

D-Glucose-6-Phosphate, Disodium Salt, Dihydrate

C6H11Na2O9P•2H2OFormula Weight: 340.13 (dihydrate)Product specifications:Form: White powderAssay (enzymatic): ≥ 95%Storage: -20°C, desiccatedCAS #3671-99-6

16185 5 gm 10 gm

Glucose-6-Phosphate Dehydrogenase

(E.C.1.1.1.49)Source: Leuconostoc mesenteroidesProduct specifications:Form: Ammonium sulfate suspensionActivity: ≥ 400 units/mg protein (NAD units)Activity: ≥ 220 units/mg protein (NADP units)Protein Concentration: ≥ 8 mg/mlContaminating Activities:

Hexokinase: ≤ 0.01%Phosphoglucose Isomerase: ≤ 0.005%Phosphogluconate Dehydrogenase: ≤ 0.001%Creatine Kinase: ≤ 0.001%Glutathione Reductase: ≤ 0.001%Phosphoglucomutase: ≤ 0.001%Myokinase: ≤ 0.001%Lactate Dehydrogenase: ≤ 0.01%

Note: The above product is available in manufac-turing quantities as a lyophilized powder or an ammonium sulfate suspension. Please inquire for bulk pricing.

Storage: 4°C

16171 1,000 units 5,000 units 10,000 units

Biochemicals


Glucose-6-Phosphate Dehydrogenase

(E.C.1.1.1.49)Source: YeastProduct specifications:Form: Ammonium sulfate suspensionActivity: ≥ 250 units/mg proteinUnit Definition: One unit reduces 1 µmol NADP/

minute at 25°C, pH 7.8.Contaminating Activities:

Glutathione Reductase: ≤ 0.2%Hexokinase: ≤ 0.02%Phosphogluconate Dehydrogenase: ≤ 0.01%Glucosephosphate Isomerase: ≤ 0.01%Phosphoglucomutase: ≤ 0.01%Creatine Kinase: ≤ 0.001%Myokinase: ≤ 0.01%ATPase: ≤ 0.001%

Note: The above product is available in manufac-turing quantities as a lyophilized powder or an ammonium sulfate suspension. Please inquire for bulk pricing.

Storage: 4°C

16176 1,000 units 5,000 units 10,000 units

Glucose-6-Phosphate Glutamate Dehydrogenase

(E.C.1.4.1.3)Source: Bovine LiverProduct specifications:Form: Lyophilized powderActivity: ≥ 10 units/mg powder, ~ 40 units/mg

enzyme proteinContaminating Activity:

Malate dehydrogenase: < 0.01%Unit Definition: One unit catalyzes the oxidation of

1 µmol of 2-oxoglutarate/minute at 25°C under assay conditions not containing ADP.

Storage: -20°C

16250 100 mg 500 mg 1 gm

L-Glutamic Acid(L-α-Aminoglutaric Acid)

HOOCCH2CH2CH(NH2)COOHFormula Weight: 147.13Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity: By IR[α]20

D : +31.2° to +32.2° (c=10 in 2N HCl)Loss on Drying: ≤ 0.1%Heavy Metals (Pb): ≤ 20 ppmArsenic (As): ≤ 1.5 ppmLead (Pb): ≤ 10 ppmCAS #56-86-0

16215 1 kg 5 kg 25 kg

L-Glutamic Acid, Monosodium Salt, Monohydrate(Monosodium Glutamate)

USBioAnalyzedC5H8NNaO4

•H2OFormula Weight: 187.13Product specifications:Form: White crystalline powderAssay: 99.0 - 100.5%Identity: By IRIdentification (A, B, C,): Passes testsClarity and color of solution: Passes testpH (5%, H2O): 6.7 - 7.2[α]20

D : +24.8° to +25.3° (c=10 in 2N HCl)Loss on Drying (100°C, 5 hours): ≤ 0.5%Heavy Metals (Pb): ≤ 20 ppmChloride (Cl): ≤ 0.25%Lead (Pb): ≤ 10 ppmResidual Solvents: Meets requirementsCAS #6106-04-3

16245 5 kg

L-Glutamine(L-α-Aminoglutamic Acid)

H2NCOCH2CH2CH(NH2)COOHFormula Weight: 146.14Product specifications:Form: White crystalline powderAssay: ≥ 98.5%Identity: By IR[α]20

D : +6.3° to +7.3° (c=4 in H2O)Loss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 10 ppmArsenic (As): ≤ 3 ppmChloride (Cl): ≤ 0.021%Iron (Fe): ≤ 10 ppmSulfate (SO4): ≤ 0.028%CAS #56-85-9

16285 100 gm 250 gm 500 gm 1 kg

Glutamyl Cysteinyl Glycine

See Glutathione, page 218

Glutathione-Oxidized

C20H32O12N6S2•H2O

Formula Weight: 612.63Product specifications:Form: White lyophilized powderAssay (HPLC): ≥ 95%Identity: By IR[α]20

D : -92° to -106° (c=4 in H2O)Moisture (KF): ≤ 8%Heavy Metals (Pb): ≤ 10 ppmIron (Fe): ≤ 10 ppmStorage: 4°C, desiccatedCAS #27025-41-8

16320 250 mg 1 gm

Glutathione-Reduced(γ-L-Glutamyl-Cysteinyl Glycine)

C10H17O6N3SFormula Weight: 307.32Product specifications:Form: White crystalline powderAssay: ≥ 98%[α]20

D : -15° to -18° (c=2 in H2O)Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.2%Heavy Metals (Pb): ≤ 20 ppmArsenic (As): ≤ 2 ppmIron (Fe): ≤ 5 ppmCAS #70-18-8

16315 25 gm 100 gm

Biochemicals


Glycerin

See Glycerol, page 219

Glycerol(Glycerin)

ULTRAPURE

MB GradeHOCH2CH(OH)CH2OHFormula Weight: 92.09Product specifications:Form: Clear, colorless, viscous liquidAssay (LISP): 99.0 - 101.0%Identity: By IRSpecific Gravity (25°C): ≥1.249Color (APHA): ≤ 10Neutrality: Passes testResidue on Ignition: ≤ 0.007%Moisture: ≤ 0.5%Fatty Acid Esters: ≤ 0.3 ml of 0.5N NGOHSilver Reducing Substances: Passes testReadily Carbonizable Substances: Passes testRefractive Index: 1.470-1.475Chlorinated Compounds: ≤ 0.003%Chloride (Cl): ≤ 0.001%Sulfate (SO4): ≤ 0.002%Arsenic (As): ≤ 1.5 ppmCalcium (Ca): ≤ 5 ppmIron (Fe): ≤ 5 ppmMagnesium (Mg): ≤ 1 ppmMercury (Hg): ≤ 10 ppmZinc (Zn): ≤ 2 ppmHeavy Metals (Pb): ≤ 2 ppmDNase (endo): Passes testDNase (exo): Passes testRNase: Passes testProtease: Passes testApplications:� Used for long-term storage of bacterial cultures.� Step gradients for bacteriophage/particle purifica-

tion� Mammalian cell transfection� Storage buffers for enzymesTypical storage for bacterial cultures: Saturated bacterial culture 0.85 ml Glycerol, Ultrapure, sterile 0.15 ml Vortex the solution and transfer to a screw cap

storage tube. Freeze quickly and store at -70°C.CAS #56-81-5

16374 500 ml 1 L

Glycerol Tolerant Gel (GTG) Buffer, 20X SolutionULTRAPURE

MB GradeGlycerol Tolerant Gel (GTG) Buffer is formulated to resolve the problem of gel distortion associated with sequencing samples that contain glycerol. Standard TBE buffer causes gel distortion between 300- 500 bases as a result of reactions between glycerol and boric acid to form an anionic ester compound, which migrates in the sequencing gel. This buffer also works equally well with samples that contain no glycerol.Form: Solution consists of 1.78 M Tris, 0.57 M

Taurine, and 0.01 M EDTA. Solution is 0.2 µm fil-tered and dispensed into durable, square Nalgene bottles. Dilute to 1X with high purity dH2O for a solution consisting of 0.089 M Tris, 0.029 M Taurine, and 0.5 mM EDTA. Total volume is 1.5 liters of 20X buffer or 30 liters of 1X buffer.

pH (1:20, 25°C): 8.9 ± 0.1Conductivity 1:20, 830 ± 100 µmhos/cmTesting: Meets or exceeds functional test criteria for

high quality gel electrophoresis. Application: � Specifically formulated to alleviate the effects of

glycerol in DNA sequencing or PAGE gels.Storage: Ambient. Crystals which form in the cold at

low temperatures can be redissolved by warming briefly.

75827 1 L

β-Glycerophosphate, Sodium Salt, Pentahydrate

C3H5(OH)2PO4Na2•5H2O

Formula Weight: 306.11Alpha Content: 50%CAS #1555-56-2

21655 100 gm 500 gm 1 kg

GlycineULTRAPURE

MB GradeH2NCH2CO2HFormula Weight: 75.07Product specifications:Form: White powderAssay: ≥ 99.0%Identity: By IRλmax: 260 nm, 1 M, 1 cm: ≤ 0.05 280 nm, 1 M, 1 cm: ≤ 0.05Loss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.1%Readily Carbonizable Substances: Passes testHydrolyzable Substances: Passes testArsenic (As): ≤ 1 ppmBarium (Ba): ≤ 1 ppmCadmium (Cd): ≤ 1 ppmCalcium (Ca): ≤ 5 ppmChloride (Cl): ≤ 50 ppmCopper (Cu): ≤ 1 ppmIron (Fe): ≤ 1 ppmLead (Pb): ≤ 1 ppmMagnesium (Mg): ≤ 1 ppmManganese (Mn): ≤ 1 ppmSulfate (SO4): ≤ 50 ppmZinc (Zn): ≤ 1 ppmContaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testCAS #56-40-6

16407 500 gm 1 kg 5 kg

Turn to us as your single sourcingsolution for raw materials. Bulk quantities are available.

Biochemicals


Glycine

ACS Reagent GradeH2NCH2COOHFormula Weight: 75.07Product specifications:Form: White crystalline powderAssay: ≥ 98.5%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 0.002%Chloride (Cl): ≤ 0.005%Sulfate (SO4): ≤ 0.005%Ammonium (NH4): ≤ 0.005%Substances Darkened by Sulfuric Acid: Passes testHydrolyzable Substances: Passes testCAS #56-40-6

16372 2.5 kg

Glycine

USBioAnalyzedH2NCH2CO2HFormula Weight: 75.07Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity: By IRLoss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.1%Sulfate (SO4): ≤ 65 ppmHeavy Metals (Pb): ≤ 20 ppmChloride (Cl): ≤ 70 ppmHydrolyzable Substances: Passes testResidual Solvents: Meets requirementsCAS #56-40-6

16405 1 kg 5 kg 25 kg

GlycogenULTRAPURE

Source: OystersProduct specifications:Form: Light tan powderIdentity: By IR[α]20

D : +170° to 200° (c=0.2, H2O, dry basis)Solubility (0.25%, 1 M NaCl): Passes testLoss on Drying (110°C, 4 hours.): ≤ 15%CAS #9005-79-2

16445 5 gm 10 gm 25 gm

Glycogen, 20 mg/mlULTRAPURE

MB GradeSource: OystersProduct specifications:Concentration: 20 mg/ml in nuclease-free waterNote: Glycogen is an inert carrier which significantly

increases the efficiency of extracted DNA and RNA by ethanol precipitation.

Contaminant Testing:DNase (endo): Passes testDNase (exo): Passes testRNase: Passes test

Storage: -20°C

77534 1 ml 5 x 1 ml

Glycyl-Glycine

NH2CH2CONHCH2COOHFormula Weight: 132.12Product specifications:Form: White crystalline powderAssay: ≥ 98.5%Chromatography (TLC): HomogeneousIdentity: By IRpKa (25°C): 8.25 ± 0.15Solubility (10%, H2O): Clear and colorlessLoss on Drying: ≤ 0.2%Chloride (Cl): ≤ 0.02%CAS #556-50-3

16505 100 gm 1 kg

GTP

See Guanosine-5’-Triphosphate, page 221

Guanidine HydrochlorideULTRAPURE

MB Grade(NH2)2C=NH•HClFormula Weight: 95.53Product specifications:Form: White/clear crystalsIdentity: By IRAssay: ≥ 99.5%Solubility (6 M, H2O): Clear and colorlessA (225 nm, 6 M, 1 cm): ≤ 0.2A (260 nm, 6 M, 1 cm): ≤ 0.03Melting Point: 184 ± 3°CResidue on Ignition: ≤ 0.05%Aldehyde: None detectedAmmeline: None detectedArsenic (As): ≤ 1 ppmCopper (Cu): ≤ 1 ppmIron (Fe): ≤ 2 ppmLead (Pb): ≤ 5 ppmSulfate (SO4): ≤ 50 ppmZinc (Zn): ≤ 1 ppmDNase (endo): Passes testRNase: Passes testProtease: Passes testApplications:� Used for extraction of RNA.� Preparation of high molecular weight genomic

DNA� Solubilizing inclusion bodiesPhenol/Chloroform RNA Isolation MethodA. Guanidine HCl Homogenization Buffer for RNA

Isolation: 8.0 M Guanidine HCl, 0.1 M Sodium Acetate, pH 5.5, 5.0 mM Dithiothreitol, 0.5% Sodium Lauryl Sarcosinate

B. Extraction with Phenol: Most extractions are per-formed using equal volumes of equilibrated phe-nol with respect to the sample volume. Normally, the extraction is performed twice with phenol and once with a 1:1 solution of phenol/chloro-form. Hence, it is estimated that 25 ml of phenol is required for every 10 ml of sample.

Note: If regular phenol is purchased, it will need to be equilibrated before use. For this reason, many customers may prefer to purchase phenol that is already equilibrated (PN 75829).

C. RNA Resuspension Buffer: TE, pH 7.5.CAS #50-01-1

75823 100 gm 500 gm

Biochemicals


Guanidine Hydrochloride

(NH2)2C=NH•HClFormula Weight: 95.53Product specifications:Form: White/clear crystalsIdentity: By IRMelting Point: 184 - 190°CLoss on Drying: ≤ 0.50%Note: Contains some water-insoluble matter.CAS #50-01-1

34007 1 kg 5 kg

Guanidine ThiocyanateULTRAPURE

(NH2)2C=NH•HNCSFormula Weight: 118.16Product specifications:Form: White crystalline powderAssay: ≥ 99%Identity: By IRA (280 nm, 6 M, 1 cm): ≤ 0.8Solubility (5%, H2O): Passes testSolubility (5%, MeOH): Passes testMelting Point: 120 ± 2°CMoisture (KF): ≤ 1%Copper (Cu): ≤ 0.5 ppmIron (Fe): ≤ 5 ppmLead (Pb): ≤ 1 ppmZinc (Zn): ≤ 2 ppmApplications:� Used to extract RNA from cells and tissue.� Denatures protein, thus inactivating ribonu-

cleases.Solution for Guanidine Thiocyanate RNA Extraction: Guanidine Thiocyanate 4.0 M Dithiothreitol 0.1 mM Sodium Acetate, pH 5.2 20 mM N-Lauryl sarcosine 0.5% Heat slightly (65°C) to dissolve the guanidine

thiocyanate and sodium acetate. Add the dithio-threitol and N-lauryl sarcosine. The pH should be about 5.5. Filter and store at room temperature.

Cesium Chloride RNA Isolation MethodA. Guanidine Thiocyanate Homogenization Buffer

for RNA Isolation: 4.0 M Guanidine Thiocyanate, 0.1 M Tris, pH 7.5, 1.0% 2-mercaptoethanol.

B. Cesium Chloride RNA Isolation Solution: Most researchers layer their RNA homogenates onto a cushion of 5.7 M Cesium Chloride.

C. RNA Resuspension Buffer: TE, pH 7.5.CAS #593-84-0

75818 100 gm 250 gm 1 kg

Guanosine-5’-Diphosphate, Disodium Salt(GDP)


Formula Weight: 541.21Reference constants:λmax (pH 7, NRC): 253 nmεmax (pH 7, NRC): 13.7 x 103

ε260 (pH 7, DBR): 11.8 x 103


16785 100 mg 500 mg 1 gm

Guanosine-5’-Triphosphate, Sodium Salt, 100 mM Solution(GTP)

ULTRAPURE

C10H12N5O14P3Na4


77243 25 µmol (250 µl)

Guanosine-5’-Triphosphate, Disodium Salt(GTP)

C10H14N5O14P3Na2

Formula Weight: 567.14Product specifications:Assay: ≥ 95%λmax: 252 nm ± 1nmεmax: (13.7 ± 0.7) x103

Spectral Ratios (pH 7): A250/A260: 1.15 ± 0.03 A280/A260: 0.65 ± 0.03Moisture (KF): ≤ 10.0%Reference constants:λmax (pH 7, NRC): 252 nmεmax (pH 7, NRC): 13.7 x 103

ε260 (pH 7, DBR): 11.7 x 103

Storage: -20°C in tightly closed containers.CAS #56001-37-7

16800 25 mg 100 mg 500 mg 1 gm

Hemin

Source: PorcineC34H32O4N4FeCl Formula Weight: 651.94 Product specifications:Form: Dark blue/purple to black crystalline powderAssay: ≥ 96%Identity: By IRMoisture (KF): ≤ 2.0%Solubility (0.02%, 0.1N NaoH): Passes testCAS #16009-13-5

16880 10 gm 50 gm

Heparin, Sodium Salt

USBioAnalyzedProduct specifications:Form: White to pale cream colored powder.Assay: ≥ 180 USP Heparin units/mg (dry basis)Identification A: Passes testIdentification B: Passes testIdentification C: 0.9 - 1.1Identification D: Passes testAnti-Factor Xa Activity: Record USP U/mgAnti-Factor IIa Potency (as is): Record USP U/mgResidue on Ignition: 28.0 to 41.0%Nitrogen Content: 1.3 to 2.5% (dry basis)Heavy Metals: ≤ 0.003% Organic Impurities Limit of Galactosamine: ≤ 1%Nucleotidic Impurities: ≤ 0.20Absence of Oversulfated Chondroitin Sulfate A:

Passes testAbsence of Oversulfated Chondroitin Sulfate B:

Passes testProtein Impurities: ≤ 1.0%pH (1%, H2O): 5.0 to 7.5Bacterial Endotoxins: ≤ 0.03 Endotoxin units/

Heparin unitLoss on Drying: ≤ 5.0% Residual Solvents: Passes TestStorage: Ambient, in tightly closed containers.CAS #9041-08-1

16920 50,000 units 100,000 units 500,000 units 1 mu

Biochemicals


HEPES, 1 M Solution, pH 7.3ULTRAPURE

MB GradeForm: 1 M solution of HEPES in high purity H2O, pH

7.3 (at 37°C). Solution is 0.2 µm filtered and dis-pensed into durable, square Nalgene bottles.

pH (37°C): 7.3 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testStorage: Ambient, store at +4°C after opening.

16924 100 ml 500 ml 1 L

HEPES(N-[2-Hydroxyethyl]piperazine-N’ [2-Ethane-sulfonic Acid])

ULTRAPURE

C8H18N2O4SFormula Weight: 238.30Product specifications:Form: White crystalline powderAssay: ≥ 99.0% (dry basis)Identity: By IRpKa (20°C): 7.55 ± 0.15A (250 nm, 1 M, 1 cm): ≤ 0.2Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Chloride (Cl): ≤ 0.005%Phosphate (PO4): ≤ 0.001%Sulfate (SO4): ≤ 0.01%Iron (Fe): ≤ 5 ppmHeavy Metals (Pb): ≤ 10 ppmCAS #7365-45-9

16926 100 gm 250 gm 1 kg 5 kg

HEPES, Sodium Salt

HO(CH2)2(C4H8N2)(CH2)2SO3NaFormula Weight: 260.29Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (dry basis)Identity: By IRpKa (20°C): 7.55 ± 0.15pH (0.1 M, H2O): 10.2 ± 0.8A (250 nm, 5%, 1 cm): ≤ 0.30A (260 nm, 5%, 1 cm): ≤ 0.175Solubility (5%, H2O): Clear solutionLoss on Drying: ≤ 4.0%Heavy Metals (Pb): ≤ 10 ppmStorage: Hygroscopic, store in tightly closed con-

tainer.CAS #75277-39-3

16928 100 gm 1 kg

Hexokinase

(E.C.2.7.1.1)Source: YeastProduct specifications:Form: Lyophilized, ammonium sulfate-free powderActivity: ≥ 140 units/mg protein ≥ 80 units/mg powderUnit Definition: One unit converts 1 µmol of glucose

to glucose-6-phosphate per minutes at 25°C, pH 7.5 in the presence of ATP.

Contaminating Activities: PGI: ≤ 0.1% G-6-PDH: ≤ 0.01% PGluM: ≤ 0.01% MK: ≤ 0.01% ATPase: ≤ 0.003% PGDH: ≤ 0.005% CK: ≤ 0.005% GR: ≤ 0.05%Note: The above product is available in manufac-

turing quantities as a lyophilized powder or an ammonium sulfate suspension. Please inquire for bulk pricing.


16965 1,000 units 10,000 units 25,000 units 50,000 units

L-Histidine, Free Base

USBioAnalyzed(C3N2H3)CH2CH(NH2)COOHFormula Weight: 155.15Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% C6H9N3O2 (dry basis)Identity: By IRChromatographic Purity: Passes testpH (2%, H2O): 7.0 - 8.5[α]25

D : +12.6° to +14.0° (c=11 in 6N HCl)Loss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.4%Heavy Metals (Pb): ≤ 0.0015%Chloride (Cl): ≤ 0.05%Iron (Fe): ≤ 0.003%Sulfate (SO4): ≤ 0.03%CAS #71-00-1

17070 100 gm 1 kg

L-Histidine, Monohydrochloride, Monohydrate

(C3H3N2)CH2CH(NH2)COOH•HCl•H2OFormula Weight: 209.63Product specifications:Form: White crystalline powderAssay: ≥ 98.0%Identity: By IRChromatography (TLC): Homogeneous[α]20

D : +9.2° to +10.6° (c=11 in 6N HCl)Loss on Drying: ≤ 0.3%Residue on Ignition: ≤ 0.10%Heavy Metals (Pb): ≤ 10 ppmAmmonium (NH4): ≤ 0.02%Arsenic (As2O3): ≤ 1 ppmIron (Fe): ≤ 30 ppmSulfate (SO4): ≤ 0.03%CAS #5934-29-2

17080 100 gm 1 kg 5 kg

L-Hydroxysuccinic Acid

See L-Malic Acid, page 227

Biochemicals


ImidazoleULTRAPURE

C3H4N2

Formula Weight: 68.08Product specifications:Form: White to cream crystalline flakes/powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (25°C): 6.95 ± 0.15Melting Point: 88 - 92°CMoisture: ≤ 1.0%Iron (Fe): ≤ 30 ppmHeavy Metals (Pb): ≤ 10 ppmCAS #288-32-4

17525 100 gm 500 gm 1 kg 5 kg

IPTG

See Isopropyl-β-D-Thiogalactopyranoside, page 223

Isocitrate Dehydrogenase, (NADP+ Specific), Recombinant[r-ICDH (NADP+)]

NADP+ Specific(E.C.1.1.1.42)Source: Yeast recombinant in S. cerevisiaeProduct specifications:Form: 50% Glycerol SolutionSpecific Activity: ≥ 30 units/mg proteinpH: 7.0 ± 0.5Unit Definition: One unit converts 1 µmol of isoci-

trate to α-Ketoglutarate/minute at 25°C, pH 8.5.Contaminating Activity: ICDH (NAD+): ≤ 0.5%Note: The above product is available in manufactur-

ing quantities as an ammonium sulfate suspen-sion or a glycerol solution. Please inquire for bulk pricing.

Storage: -20°C


DL-Isoleucine

CH3CH2CH(CH3)CH(NH2)COOHFormula Weight: 131.17Product specifications:Form: White crystalline powderAssay: ≥ 98.0%Identity: By IRChromatography (TLC): HomogeneousLoss on Drying: ≤ 0.3%Residue on Ignition: ≤ 0.1%Heavy Metals: ≤ 20 ppmChloride (Cl): ≤ 0.02%Iron (Fe): ≤ 50 ppmSulfate (SO4): ≤ 0.05%CAS #443-79-8

17820 100 gm 500 gm 1 kg

L-Isoleucine

USBioAnalyzedCH3CH2CH(CH3)CH(NH2)COOHFormula Weight: 131.17Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% C6H13NO2 (dry basis)Identity: By IRChromatographic Purity: Passes testpH (1%, H2O): 5.5 - 7.0[α]20

D : +38.9° to +41.8° (c=4 in 6N HCl, 20°C)Loss on Drying: ≤ 0.3%Residue on Ignition: ≤ 0.3%Sulfate (SO4): ≤ 0.03%Heavy Metals (Pb): ≤ 0.0015%Chloride (Cl): ≤ 0.05%Iron (Fe): ≤ 0.003%CAS #73-32-5

17825 10 gm 100 gm 500 gm 1 kg

Isopropyl-β-D-Thiogalactopyranoside (IPTG), Dioxane FreeULTRAPURE

C9H18O5SFormula Weight: 238.30Product specifications:Form: White to off-white powderAssay: ≥ 99%Identity: By IR[α]20

D : -31.5°C ± 1° (c=1 in H2O)Melting Point: 109 - 114°CMoisture (KF): ≤ 2.0%Dioxane (GC): ≤ 0.02%Applications:� Induces the lac operon in E. coli by binding and

inactivating the lac repressor.� Selection for lac Y mutants� Selection for recombinant plasmids containing

β-galactosidase fusion proteins� Induces the cellular content of lactose permease.Storage: -20°C, desiccated, protect from light.CAS #367-93-1

17886 1 gm 5 gm 25 gm 100 gm

Kanamycin Sulfate

C18H36N4O11•H2SO4

Formula Weight: 582.58Product specifications:Form: White to off-white powderPotency:

≥ 750 µg of C18H36N4O11/mg powder (dry basis)Identity (A, B, C): Passes testspH (1%, H2O): 6.5 - 8.5Loss on Drying: ≤ 1.5%Residue on Ignition: ≤ 0.5%Specific Rotation: +112° ± 123° (c=1 in H2O)Mode of Action: Binds to ribosomal components

and inhibits protein synthesis. Kanamycin is inac-tivated by an aminoglycoside phospho transfer-ase (MW 25,000) in the periplasmic space.

Application:� A selectable marker for cosmid vectors used in

transfection of mammalian cells.Storage: 4°CCAS #25389-94-0

17924 5 gm 25 gm

Biochemicals


α-Ketoglutaric Acid, Free Acid

C5H6O5

Formula Weight: 146.10Product specifications:Form: White to off-white crystalline powderAssay: ≥ 98.0% (as is)Identity: By IRMelting Point: 114 - 119°CLoss on Drying: ≤ 1.0%CAS #328-50-7

17950 100 gm 500 gm 1 kg

α-Ketoglutaric Acid, Disodium Salt, Dihydrate

C5H4O5Na2•2H2O

Formula Weight: 226.09 (dihydrate) 190.06 (anhydrous)Product specifications:Form: White powderAssay: ≥ 98.0% (as is)Identity: By IRMoisture: 14.0 - 18.0%CAS #305-72-6

17955 5 gm 100 gm 1 kg

L- (+)-Lactic Acid, Lithium Salt(Lithium Lactate)

C3H5LiO3

Formula Weight: 96.01Product specifications:Form: White crystalline powder[α]20

D : -11.5° ± 1.5° (c=10, H2O)Solubility: Passes testLoss on Drying: ≤ 0.5%CAS #27848-80-2

18160 100 gm

Lactoferrin

Form: Off-white powderPurity: ≥ 95%Protein (as is): ≥ 93%Ash: ≤ 1.5%Storage: 4°C, in tightly closed containers.

18177 100 mg 500 mg 1 gm

D-(+)-Lactose, Monohydrate

USBioAnalyzedC12H22O11

•H2OFormula Weight: 360.31Product specifications:Form: White powderClarity and Color of Solution: Passes testIdentity (A, B, C): Passes tests[α]20

D : +54.4° to +55.9° (c=10 in 0.2 ml 6N NH4OH/100 ml H2O)

Acidity or Alkalinity: Passes testLoss on Drying: ≤ 0.5%Moisture (KF): 4.5 - 5.5%Residue on Ignition: ≤ 0.1%Protein and Light-Absorbing Impurities: Passes testHeavy Metals (Pb): ≤ 5 ppmMicrobial Limits: Passes testCAS #64044-51-5

18185 5 kg

N-Lauroyl Sarcosine, Sodium Salt(Sodium-N-Lauryl Sarcosine)

ULTRAPURE

HO2CCH2CH3NCO(CH2)10CH2•Na

Formula Weight: 293.38Product specifications:Form: White to faint white-yellow powderIdentity: By IRMoisture (KF): ≤ 5.0%pH (10%, H2O): 7.5 - 8.5Solubility (10%, H2O): Clear to slightly hazy, color-

less to faint yellow solutionCAS #137-16-6

21653 50 gm 100 gm

Lauryl Sulfate, Lithium Salt


Lauryl Sulfate, Sodium Salt


LB Agar, Ready-Made Powder

Form: Off-white free flowing powder.pH (3.5%, H2O): 7.0 ± 0.2Contents: Casein Peptone: 10 gm/L Yeast Extract: 5 gm/L Sodium Chloride: 5 gm/L Agar: 15 gm/LTo use: Dissolve 35 gm powder per liter of purified

water. Autoclave, cool and make plates.Application: � For the propagation of bacteria

75851 250 gm 1 kg

LB Broth, Ready-Made Powder

Form: Luria-Bertani Broth. Off-white free flowing powder.

pH (2.0%, H2O): 7.0 ± 0.2Contents: Casein Peptone: 50% Yeast Extract: 25% Sodium Chloride: 25%To use: Dissolve 20 gm powder per liter of purified

water. Autoclave.Application: � For the propagation of bacteria

75852 250 gm 1 kg

Lecithin

Source: SoybeanProduct specifications:Form: Off-white to light tan granular powderMoisture: ≤ 1.5% (at time of packaging)Acetone Insolubles: ≥ 95.0%Hexane Insolubles: ≤ 0.3%CAS #8002-43-5

18245 500 gm 1 kg

Leptomycin B

C33H48O6

Formula Weight: 540.73Form: Leptomycin B is an antifungal antibiotic that

blocks CRM1 mediated nuclear export of proteins containing Nuclear Export Signal (NES). It inac-tivates CRM1 by covalently modifying the sulf-hydryl residue of Cysteine-529. This inhibits the formation of a complex between CRM1, Ran, and a protein with a NES. This results in the nuclear retention of the NES-bearing protein(1). Inhibiting the CRM1-mediated nucleo-cytoplasmic export of proteins bearing a NES produces the nuclear accumulation of a large number of proteins(2) such as Actin, c-Abl, Cdc25, Cyclin B1, HDM2, HIV-1, IΚBα, and p53.

References1. Kudo, N., Matsumori, N., Taoka, H., Fujiwara,

D., Schreiner, E. P., Wolff, B., Yoshida, M. and Horinouchi, S. (1999) Proc. Natl. Acad. Sci. USA 96, 9112-9117.

2. Lain, S., Xirodimas, D. and Lane, D. (1999) Experimental Cell Research 253, 315-324.

CAS #87081-35-4

25110 1 µg

Biochemicals


DL-Leucine

(CH3)2CHCH2CH(NH2)COOHFormula Weight: 131.17Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (as is)Identity: By IRChromatography (TLC): HomogeneousLoss on Drying: ≤ 0.3%CAS #328-39-2

18270 100 gm 1 kg

L-Leucine

C6H13NO2

Formula Weight: 131.17Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity: By IR[α]20

D : +14.5° to +16.5° (c=4 in 6N HCl dry basis)Loss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.1%Lead (Pb): ≤ 10 ppmHeavy Metals (Pb): ≤ 20 ppmCAS #61-90-5

18280 100 gm 1 kg 5 kg

LeupeptinULTRAPURE

C20H38N6O4•1⁄2 H2SO4

Formula Weight: 475.59Form: White powderApplications:� Leupeptin, a protease inhibitor, will strongly

inhibit Trypsin, Papain, Plasmin, Thrombokinase, Kallikrein, and Cathepsin B. The half-maximal inhibitory concentration ranges from 0.5 to 75 µg/ml, depending on the enzyme and the substrate.

� Leupeptin does not inhibit Chymotrypsin, Elastase, Renin, or Pepsin.

Storage: -20°CCAS #55123-66-5

18413 5 mg 25 mg 100 mg

DL-α-Lipoic Acid(DL-6,8-Thioctic Acid)

C8H14O2S2

Formula Weight: 206.33Product specifications:Form: Light yellow crystalline powderAssay: ≥ 98.0% (dry basis)Identity: Passes testLoss on Drying: ≤ 0.5%Storage: In tightly closed, light-resistant containers.CAS #1077-28-7

18505 25 gm 50 gm 100 gm

DL-α-Lipoic Acid (DL-6,8-Thioctic Acid)

C8H14O2S2

Formula Weight: 206.33Product specifications:Form: Light yellow crystalline powderAssay: ≥ 98.0% (dry basis)Identity: By IRMoisture (KF): ≤ 0.5%Melting Point: 60-62°CSolubility ( 3 gm/60 ml EtOH): Light yellow solutionStorage: Ambient, in tight, light resistant containersCAS# 1077-28-7

18522 50 gm 100 gm 1 kg

Lithium Chloride (LiCl), 7.5 M SolutionULTRAPURE

MB GradeForm: 7.5 M solution of lithium chloride (LiCl) in

high purity dH2O. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles.

Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testCAS #7447-41-8

75902 100 ml

Lithium Dodecyl Sulfate(Lithium Lauryl Sulfate)

ULTRAPURE

CH3(CH2)11OSO3LiFormula Weight: 272.33Product specifications:Assay (acidity): ≥ 98%Assay (C12 by GC): ≥ 98%A (230 nm, 3%, 1 cm): ≤ 0.2A (280 nm, 3%, 1 cm): ≤ 0.1Loss on Drying: ≤ 1%Chloride (Cl): ≤ 0.01%Copper (Cu): ≤ 5 ppmIron (Fe): ≤ 1 ppmLead (Pb): ≤ 5 ppmCAS #2044-56-6

32816 25 gm 100 gm

Lithium Lactate

See L-(+)-Lactic Acid, Lithium Salt, page 224

Lithium Lauryl Sulfate


Luria Agar, Ready-Made Powder

Form: Off-white free flowing powderpH (4.0%, H2O): 7.0 ± 0.2Contents: Casein Peptone: 25% Yeast Extract: 12.5% Sodium Chloride: 2.5% Agar: 37.5%To use: Dissolve 40 gm powder per liter of purified

water. Autoclave, cool and make plates.Application: � For the propagation of bacteria

75853 250 gm 1 kg

Luria Broth, Ready-Made Powder

Form: Off-white free flowing powderpH (2.5%, H2O): 7.0 ± 0.2Contents: Casein Peptone: 40% Yeast Extract: 20% Sodium Chloride: 40%To use: Dissolve 25 gm powder per liter of purified

water. Autoclave.Application: � For the propagation of bacteria

75854 250 gm 1 kg

Biochemicals


L-Lysine, Hydrochloride

NH2(CH2)4CH(NH2)COOH•HClFormula Weight: 182.65Product specifications:Form: White crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IR[α]20

D : +19.0 to +21.5° (c=8 in 6N HCl)Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): < 10 ppmAmmonium (NH4): ≤ 0.02%Arsenic (As2O3): ≤ 1 ppmIron (Fe): ≤ 20 ppmSulfate (SO4): ≤ 0.03%CAS #657-27-2

18585 1 kg 5 kg 10 kg

LysozymeULTRAPURE

(E.C.3.2.1.17)Source: Chicken Egg WhiteProduct specifications:Form: Lyophilized powderActivity: Approximately 20,000 units/mgUnit Definition: One unit causes a decrease in

absorbance of 0.001 per minute at 450 nm, 25°C and pH 7.0 with Micrococcus lysodeikticus as a substrate.

Moisture: ≤ 6.0%Chloride: 3.2 - 4.2%pH (2%, H2O): 3.0 - 4.0Storage: -20°C, desiccated

18645 5 gm 10 gm 25 gm 100 gm

M13mp18, Single-Stranded DNA

See Molecular Biology Products & Kits section, page 129

Magnesium Chloride (MgCl2), 1 M SolutionULTRAPURE

MB GradeForm: 1 M solution of magnesium chloride (MgCl2)

in high purity dH2O. Solution is 0.2 µm filtered and dispensed into vials and/or durable, square Nalgene bottles.

Assay: 1.00 M ± 0.01 M (EDTA titration)Contaminant Testing: Heavy Metals(Pb): ≤ 5 ppm DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes test

78641 10 x 1 ml 100 ml

Magnesium Chloride, HexahydrateULTRAPURE

ACS Reagent GradeMgCl2•6H2OFormula Weight: 203.30Product specifications:Form: White crystalline powderAssay: 99.0 - 102.0%Insoluble Matter: ≤ 0.005%Nitrate (NO3): ≤ 0.001%Phosphate (PO4): ≤ 5 ppmSulfate (SO4): ≤ 0.002%Ammonium (NH4): ≤ 0.002%Barium (Ba): ≤ 0.005%Calcium: ≤ 0.01%Manganese (Mn): ≤ 5 ppmPotassium (K): ≤ 0.005%Sodium (Na): ≤ 0.005%Strontium (Sr): ≤ 0.005%Heavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 5 ppmCAS #7791-18-6

18641 500 gm 1 kg

Malate Dehydrogenase, Lyophilized

(E.C.1.1.1.37)Source: Porcine Heart (Mitochondrial)Product specifications:Form: Lyophilized powder, ammonium sulfate freeActivity: ≥ 1,100 units/mg protein.Protein: 10 - 50%Unit Definition: One unit causes the oxidation of

1 µmol of NADH/minute at 25°C, pH 7.4.Contaminating Activities:

Fumarase (L-Malate): ≤ 0.01%LDH: ≤ 0.01%GOT: ≤ 0.01%GDH (NAD): ≤ 0.002%NADH Oxidase: ≤ 0.001%

Note: The above product is available in manufactur-ing quantities as a lyophilized powder, an ammo-nium sulfate suspension or a glycerol solution. Please inquire for bulk pricing.

Storage: -20°CCAS #9001-64-3

18670 25,000 units 100,000 units

Maleic Acid(Toxilic Acid)

Reagent GradeC4H4O4

Formula Weight: 116.07Product specifications:Form: White crystalline powderAssay: 99.0 - 101.0% (dry basis)Identity: By IRSolubility (50%, H2O): Clear and colorlessMoisture (KF): ≤ 2.0%Iron (Fe): ≤ 5 ppmHeavy Metals (Pb): ≤ 10 ppmCAS #110-16-7

18685 1 kg

DL-Malic Acid

HOOCCH(OH)CH2COOHFormula Weight: 134.09Product specifications:Form: White granular powderAssay: ≥ 98% (as is)Identity: By IRMoisture (KF): ≤ 1.0%CAS #617-48-1

18695 1 kg

Biochemicals


L-Malic Acid

C4H6O5

Formula Weight: 134.09Product specifications:Form: White crystalline powderAssay (GC): ≥ 99%Identity: By IR[α]20

D : -26.5° to -30.0° (c=5.5 in pyridine)Melting Point: 102 - 106°CMoisture (KF): ≤ 1.0%CAS #97-67-6

18700 500 gm 1 kg

Maltose, Monohydrate

High Purity GradeC12H22O11

•H2OFormula Weight: 360.31Product specifications:Form: White crystalline powderAssay (HPLC): ≥ 98%Identity: By IR[α]20

D : +130.4° ± 1.3° (c=4 in H2O, NH4OH)Solubility (10%, H2O): Passes testLoss on Drying: ≤ 6%Residue on Ignition: ≤ 0.05%Dextrins: Passes testArsenic (As): ≤ 0.5 ppmIron (Fe): ≤ 5 ppmHeavy Metals (Cu): ≤ 10 ppmCAS #6363-53-7

18724 500 gm 1 kg

D-(+)-Maltose, Monohydrate

C12H22O11•H2O

Formula Weight: 360.31Product specifications:Form: White to off-white powderMaltose Content: ≥ 92.0% (dry basis)Glucose Content: ≤ 3.0% (dry basis)Loss on Drying: ≤ 7.0%Heavy Metals (Pb): ≤ 5 ppmCAS #6363-53-7

18725 1 kg 5 kg

Mannitol


Formula Weight: 182.17Product specifications:Form: White crystalline powderAssay: 96.0 - 101.5% (dry basis)Identity: Passes test[α]25

D : +137° to +145° (c=1 in USP prep.)Melting Point: 164 - 169°CLoss on Drying: ≤ 0.3%Acidity: Passes testReducing Sugars: Passes testChloride (Cl): ≤ 0.007%Sulfate (SO4): ≤ 0.01%Arsenic (As): ≤ 1 ppmCAS #69-65-8

18755 500 gm 1 kg 5 kg

D-(+)-Mannose

C6H12O6


D : +14.2° ± 0.7° (c=4 in H2O, NH3)Loss on Drying: ≤ 0.5%CAS #3458-28-4

18775 100 gm 500 gm

MES, Anhydrous(2-(N-Morpholino) Ethane Sulfonic Acid)

C6H13NO4SFormula Weight: 195.24CAS #4432-31-9

18888 100 gm 250 gm 1 kg

MES, Monohydrate(2-(N-Morpholino) Ethane Sulfonic Acid)

ULTRAPURE

HO3S(CH2)2(C4H8NO)•H2OFormula Weight: 213.25Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (20°C): 6.15 ± 0.15A (250 nm, 1 M, 1 cm): ≤ 0.1Moisture (KF): ≤ 11%Iron (Fe): ≤ 30 ppmLead (Pb): ≤ 10 ppmSolubility (1 M, H2O, 100 ml): Clear and colorlessNote: This product varies up to a monohydrate.CAS #145224-94-8

18886 100 gm 1 kg

MES, Sodium Salt(2-(N-Morpholino) Ethane Sulfonic Acid, Sodium Salt)

C6H12NO4SNaFormula Weight: 217.22Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (20°C): 6.15 ± 0.15A (260 nm, 1 M, 1 cm): ≤ 0.1Moisture (KF): ≤ 1%Iron (Fe): ≤ 30 ppmLead (Pb): ≤ 10 ppmSolubility (1 M, H2O, 100 ml): Clear and colorlessCAS #71119-23-8

18893 100 gm 250 gm 1 kg

Visit usb.affymetrix.com for new products, protocols, and MSDS documents.

Biochemicals


Methylene Blue, Trihydrate(Methylthionine Chloride)

(C.I.52015)C16H18ClN3S•3H2OFormula Weight: 373.90Product specifications:Form: Dark green crystalline powderAssay: 98.0 - 103.0% (dry basis)Identity: By IRλmax: 664 nm ± 1Solubility (1%, H2O): Passes testpH (1%, H2O): 3.6 - 4.5Moisture (KF): 8.0 - 18.0%Application:� Used in staining RNA that is bound to nylon

and nitrocellulose membranes. Methylene blue is the stain of choice in visualizing filter bound RNA over a pre-transfer gel stain with ethidium bromide. Ethidium bromide saturates the nucleic acids and reduces the efficiency of RNA transfer.

Typical procedure:1. Soak the dried filter in 5% acetic acid at room

temperature for 15 minutes.2. Place the filter in 0.5 M sodium acetate (pH 5.2)

and 0.04% methylene blue for 5-10 minutes at room temperature.

3. Rinse the filter in water for 5-10 minutes.CAS #99-76-3

19220 100 gm 500 gm

N,N’-Methylene-bis-Acrylamide(bis-Acrylamide)

ULTRAPURE

MB Grade(CH2=CHCONH)2CH2

Formula Weight: 154.17Product specifications:Form: White crystalline powderAssay: ≥ 99.9%Identity: By IRA (290 nm, 1%, 1 cm): ≤ 0.2pH (1%, H2O): ≥ 5.5Acrylic acid: ≤ 0.02%Lead (Pb): ≤ 1 ppmGel Performance: Passes testApplication:� Cross-linking agent used in forming polyacryla-

mide gels. The porosity of the gel is determined by the chain length and the degree of cross-linking. Most sequencing gels use a ratio of 1 molecule of cross-linker for every 19 monomers of acrylamide.

CAS #110-26-9

75821 25 gm 50 gm 100 gm 1 kg

5-Methyl-2’-Deoxycytidine-5’-Monophosphate, Disodium Salt(5-Methyl dCMP)

C10H14N3O7PNa2

Formula Weight: 365.19 (anhydrous)Product specifications:Form: White powderAssay (HPLC, as nucleotide): ≥ 98%Assay (HPLC, as nucleoside): ≥ 98%Moisture (KF): ≤ 16%Phosphorylated Nucleosides:≤ 0.20% dCMP≤ 0.049% for any other phosphorylated nucleosideStorage: -20°C, protect from moisture.

19184 100 mg

4-Methylumbelliferyl-α-L-Iduronide, Sodium Salt

C16H15NaO9

Formula Weight: 374.27Storage: -20°C

33230 5 mg

Mineral OilULTRAPURE

MB GradeProduct specifications:Form: Clear, colorless oilIdentity: By IRSpecific Gravity (25°C): 0.818 to 0.880Viscosity (40°C): ≤ 33.5 centistokesNeutrality: Passes testReadily Carbonizable Substances: Passes testLimit of Polynuclear Compounds: Passes testSolid Paraffin: Passes testUse Test: Functionally tested for use in PCR.CAS #8042-47-5

71600 10 ml 25 ml 1 L

Monosodium Glutamate

See Glutamic Acid, Monosodium Salt, page 218

MOPS(3-[N-Morpholino]propanesulfonic Acid)

ULTRAPURE

HO3S(CH2)3(C4H8NO)Formula Weight: 209.26Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (20°C): 7.20 ± 0.15A (250 nm, 1 M, 1 cm): ≤ 0.1Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Chloride (Cl): ≤ 0.01%Sulfate (SO4): ≤ 0.02%Iron (Fe): ≤ 5 ppmLead (Pb): ≤ 10 ppmCAS #1132-61-2

19256 100 gm 1 kg

MTT(3-[4,5-Dimethyl-2-Thiazolyl]-2,5-Diphenyl Tetrazolium Bromide; Thiazolyl Blue Tetrazolium Bromide)

ULTRAPURE

C18H16BrN5SFormula Weight: 414.32Product specifications:Form: Yellow powderChromatography (TLC): HomogeneousIdentity: By IRSolubility: Passes testSensitivity: Passes testMelting Point: ≥ 192°CApplications:� Used in the quantitation of DNA synthesis.� Detects living and growing cells. MTT is cleaved

by active mitochondria to form a dark blue formazan product.

CAS #298-93-1

19265 500 mg 1 gm 5 gm 10 gm

β-NAD

See Nicotinamide Adenine Dinucleotide, page 229

Biochemicals


NADH

See β-Nicotinamide Adenine Dinucleotide, Reduced, page 230

NADP

See Nicotinamide Adenine Dinucleotide Phosphate, page 230

NADPH

See Nicotinamide Adenine Dinucleotide Phosphate, Reduced, page 230

α-Naphthyl Phosphate, Monosodium Salt

C10H8NaO4P•H2OFormula Weight: 264.15 (monohydrate)Product specifications:Form: White crystalline powderAssay: ≥ 97% (as is monohydrate)Chromatography (TLC): HomogeneousIdentity: By IRMoisture (KF): ≤ 10.0%Storage: 4°CCAS #1136-89-6

19391 25 gm

NBT

See p-Nitro Blue Tetrazolium Chloride, page 230

Neocuproine, Hydrochloride(2,9-Dimethyl-1,10-Phenanthroline HCl)

C14H12N2•HCl•nH2O

Formula Weight: 244.72•nH2OProduct specifications:Form: Off-white to pale greenish-tan powderAssay: ≥ 98.0% (dry basis)Identity: By IRChromatography (TLC): HomogeneousMoisture (KF): ≤ 10%Storage: 4°CCAS #7296-20-0

19430 10 gm 25 gm 100 gm 1 kg

Neomycin SulfateULTRAPURE


•3H2SO4

Formula Weight: 908.88Product specifications:Form: White to off-white powderActivity: ≥ 600 µg neomycin/mg powder (dry basis)Activity: ≥ 552 µg neomycin/mg powder (as is)Identity (A, B, C): Passes testspH (33 mg/ml): 5.0 - 7.5Loss on Drying: ≤ 8.0%Mode of Action: Binds to ribosomal components

and inhibits protein synthesis.Stock Solution: 10 mg/ml in waterWorking Concentration: 10 µg/mlStorage: Ambient, in tightly closed, light-resistant

containers.CAS #1405-10-3

19435 10 gm 25 gm

Neutral Red

C15H17N4ClFormula Weight: 288.78Product specifications:Form: Green-black crystalline powderIdentity: By IRVisual Transition Interval: pH 6.8 - Red pH 8.0 - AmberCAS #553-24-2

19465 25 gm 100 gm 1 kg

Niacinamide(Nicotinic Acid Amide; Nicotinamide)

USBioAnalyzedC6H6N2OFormula Weight: 122.12Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity (A, B): Passes testsMelting Point: 128 - 131°CLoss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Heavy Metals: ≤ 0.003%Readily Carbonizable Substances: Passes testResidual Solvents: Meets requirementsCAS #98-92-0

19480 1 kg 10 kg

Nicotinamide

See Niacinamide, page 229

β-Nicotinamide Adenine Dinucleotide(β-NAD; Coenzyme I; Diphosphopyridine Nucleotide; DPN)

C21H27N7O14P2•3H2O

Formula Weight: 717.47 (trihydrate) 663.43 (anhydrous)Product specifications:Form: White powderAssay (Enzymatic): 99 ± 4%Assay: 100 ± 3% (UV, 260 nm, pH 7.5)Identity: By IRε340 (pH 10.0): [6.3 ± 0.2] x 103

ε260 (pH 7.5): [18.0 ± 0.5] x 103


A280/A260: 0.21 ± 0.02Spectral Ratios (pH 10, reduced with ADH): A340/A260: 0.43 ± 0.01Moisture: ≤ 8%Note: The above product is available in manufactur-


15335 1 gm 5 gm 10 gm 25 gm 100 gm

β-Nicotinamide Adenine Dinucleotide, Lithium Salt, Dihydrate(β-NAD)

C21H26N7O14P2Li•2H2OFormula Weight: 705.39 (dihydrate) 663.43 (free acid)Free of LDH. Stable.Storage: -20°C

19438 1 gm 5 gm

Biochemicals


β-Nicotinamide Adenine Dinucleotide, Monosodium Salt(β-NAD)

C21H26N7O14P2Na•2H2OFormula Weight: 721.44 (dihydrate) 685.41 (anhydrous monosodium)Product specifications:Form: White to pale yellow powderAssay (Enzymatic): ≥ 95%Assay (UV 260 nm, pH 7.5): 100 ± 3%Spectral Ratios (pH 7.5): A250/A260: 0.83 ± 0.03 A280/A260: 0.21 ± 0.02Spectral Ratios (pH 10): A340/A260: 0.43 ± 0.01 A260/A340: 2.32 ± 0.06Moisture: ≤ 8%pH (0.4%, H2O): 5.35 - 6.35Sodium (Na): 3.0% ± 0.5%Storage: -20°C, protect from moisture.CAS #20111-18-6

19439 5 gm

β-Nicotinamide Adenine Dinucleotide-Reduced, Disodium, Trihydrate(β-NADH; Coenzyme I-Reduced; Triphosphopyridine Nucleotide-Reduced; DPNH)


Formula Weight: 763.45 (disodium, trihydrate) 665.25 (anhydrous, NADA)Product specifications:Form: White to cream colored powderAssay (Enzymatic): 99 ± 4%Assay (UV, 260 nm): 99 ± 4%ε260 (10 mM Tris): [14.4 ± 0.5] x 103

ε340 (10 mM Tris): [6.3 ± 0.2] x 103

Spectral Ratios (10 mM Tris):A250/A260 (red): 0.82 ± 0.03

A280/A260 (red): 0.23 ± 0.02 A340/A260 (red): 0.43 ± 0.01Moisture (KF): ≤ 8%Sodium (Na): 6.5 ± 1.5%Note: Do not use unbuffered water for dissolution

since it may cause decomposition of β-NADH.Storage: -20°C, protect from moisture.CAS #606-68-8

15345 1 gm 5 gm 10 gm 25 gm

β-Nicotinamide Adenine Dinucleotide Phosphate, Monosodium, Tetrahydrate(β-NADP; Coenzyme II-Oxidized; Triphosphopyridine Nucleotide-Oxidized; TPN)

C21H27N7O17P3Na•4H2OFormula Weight: 837.45 (monosodium tetrahydrate) 765.39 (anhydrous, NADP•Na)Product specifications:Form: White powderAssay (Enzymatic): ≥ 93%Spectral Ratios (pH 7.5): A250/A260: 0.83 ± 0.03

A280/A260: 0.21 ± 0.02Spectral Ratios (pH 7.5, reduced with G6PDH):

A340/A260: 0.43 ± 0.02A260/A340: 2.33 ± 0.11

ε260 (pH 7.5): [18.0 ± 0.8] x 103

ε340 (pH 7.5): [6.2 ± 0.4] x 103

Moisture: ≤ 8.0%Sodium (Na): 3.0 ± 1.5%Note: The above product is available in manufactur-


22655 100 mg 250 mg 500 mg 1 gm 5 gm

β-Nicotinamide Adenine Dinucleotide Phosphate-Reduced, Tetrasodium, Tetrahydrate(β-NADPH; Coenzyme II-Reduced; Triphosphopyridine Nucleotide-Reduced; TPNH)

C21H26N7O17P3•Na4

•4H2OFormula Weight: 905.41 (tetrasodium, tetrahydrate)Product specifications:Form: White to slightly off-white powderAssay (Enzymatic): ≥ 93%ε260 (10 mM Tris): [14.4 ± 0.7] x 103

ε340 (10 mM Tris): [6.2 ± 0.3] x 103

Spectral Ratios (10 mM Tris): A340/A260: 0.43 ± 0.01Moisture: ≤ 8.0%Sodium (Na): 10.0 ± 2.0%Note: The above product is available in manufactur-

ing quantities. Please inquire for bulk pricing.Storage: -20°CCAS #2646-71-1

22650 25 mg 100 mg 250 mg 500 mg 1 gm

p-Nitro Blue Tetrazolium Chloride(NBT)

ULTRAPURE

C40H30Cl2N10O6

Formula Weight: 817.64Product specifications:Form: Yellow crystalline powderIdentity: By IRSolubility (0.5% in H2O): Passes testSolubility (1% in MeOH): Passes testSensitivity: Passes testApplication:� Used with 5-bromo-4-chloro-3-indolyl phosphate

(BCIP) to detect antigen-antibody alkaline phos-phatase complexes. The NBT/BCIP mix is convert-ed to a dense blue compound and will develop at the site of antigen-antibody complexes.

NBT Stock Solution: 50 mg/ml in 70% dimethylformamideBCIP/NBT Developing Solution: 33 ml NBT (50 mg/ml) 5 ml AP Buffer (100 mM Tris-HCl, pH 9.5,

100 mM NaCl, 5 mM MgCl2) Add 16.8 ml BCIP stock (50 mg/ml in 100%

dimethylformamide) Protect from light and use within 1 hour.CAS #298-83-9

19535 250 mg 1 gm 5 gm 10 gm

4-Nitrophenyl-N-Acetyl-β-D-Galactosaminide(p-Nitrophenyl-2-Acetamido-2-Deoxy-β-D-Galactoside)

C14H18N2O8

Formula Weight: 342.30Product specifications:Form: White to off-white powderPurity (TLC): ≥ 98%Storage: -20°CCAS #14948-96-0

34128 100 mg 1 gm

Biochemicals


4-Nitrophenyl-α-L-Fucopyranoside

C12H15NO7

Formula Weight: 285.25Product specifications:Form: White powderIdentity: By IRMelting Point: 196 ± 2°CStorage: 4°CCAS #10231-84-2

34131 50 mg 250 mg 1 gm

4-Nitrophenyl-α-D-Galactopyranoside(p-Nitrophenyl-α-D-Galactoside)

C12H15NO8

Formula Weight: 301.25Form: Slightly off-white to yellowish crystalline

powderStorage: -20°CCAS #7493-95-0

34133 500 mg 1 gm 5 gm

4-Nitrophenyl-β-D-Glucopyranoside

C12H15NO8


D : -106° ± 5° (dry basis, c=1 in H2O)A (425 nm, 8 mg/ml, 1 cm): ≤ 0.07 (in 0.15 M AMP

buffer, pH 10.2)Moisture (KF): ≤ 10%Free p-Nitrophenol: ≤ 0.01%Storage: -20°CCAS #2492-87-7

19594 1 gm 5 gm

4-Nitrophenyl-β-D-Glucuronide

C12H13NO9

Formula Weight: 315.23Storage: 4°CCAS #10344-94-2

34136 100 mg 250 mg 1 gm

4-Nitrophenyl-α-D-Maltoside

C18H25NO13

Formula Weight: 463.39Product specifications:Form: White powderIdentity: By IRPurity (HPLC): ≥ 99%Moisture: ≤ 5.0%Storage: -20°CCAS #17400-77-0

34138 50 mg 100 mg

4-Nitrophenyl-β-D-Mannopyranoside(p-Nitrophenyl-β-D-Mannoside)

C12H15NO8

Formula Weight: 301.25Product specifications:Form: White to off-white crystalline powderIdentity: By IR[α]20

D : -105.0° ± 2° (dry basis, c=4 in H2O)Moisture (KF): ≤ 1.0%Free p-Nitrophenol: ≤ 0.01%Storage: -20°CCAS #35599-02-1

34140 100 mg 500 mg

4-Nitrophenyl-β-D-Xylopyranoside

C11H13NO7

Formula Weight: 271.22Form: White powderStorage: -20°CCAS #2001-96-9

34142 500 mg 1 gm

Nucleoside-5’-Triphosphates (ATP, CTP, GTP, UTP), Set of Four, 100 mM Solutions(NTPs)

ULTRAPURE

100 mM aqueous solutions, pH 7.5Storage: -20°C

4 x 25 µmol (250 µl) each NTP per pack.77245 1 pk

Nucleotide Solutions

See Adenosine-5’-Triphosphate (ATP), page 189See 2’-Deoxyadenosine-5’-Triphosphate (dATP),

page 210See 2’-Deoxycytidine-5’-Triphosphate (dCTP),

page 210See 2’-Deoxyguanosine-5’-Triphosphate (dGTP),

page 210See 2’-Deoxynucleoside-5’-Triphosphates (dNTP),

Set of Four, page 211See 2’-Deoxythymidine-5’-Triphosphate (dTTP),

page 211See 2’-Deoxyuridine-5’-Triphosphate (dUTP),

page 211See 2’,3’-Dideoxyadenosine-5’-Triphosphate

(ddATP), page 212See 2’,3’-Dideoxycytidine-5’-Triphosphate (ddCTP),

page 212See 2’,3’-Dideoxythymidine-5’-Triphosphate

(ddTTP), page 212See Guanosine-5’-Triphosphate (GTP), page 221See PCR-Nucleotide Mixes, page 232

NZCYM Broth, Ready-Made Powder

Form: Off-white free flowing powderpH (2.2%, H2O): 7.0 ± 0.2Contents: Casein Peptone: 10 gm/L Casamino Acids: 1 gm/L Magnesium Sulfate: 0.98 gm/L Yeast Extract: 5 gm/L Sodium Chloride: 5 gm/LTo use: Dissolve 22 gm powder per liter of purified

water. Autoclave.Application: � For the propagation and maintenance of lambda

bacteriophage

75855 1 kg

Oligo (dT)12-18 Primer, MW ≅ 4500

MB GradeForm: Supplied in aqueous solutionConcentration: 25 µM ± 2.5 µM2.5 nmol per tube.Storage: -20°C

77405 100 µl (2.5 nmol)

Biochemicals


Oligo p(dT)12-18 Sodium Salt(Oligo dT)

LyophilizedForm: This oligodeoxythymidylate mixture is synthe-

sized using O-cyanoethyl phosphoramidite chem-istry. After the removal of the protecting groups, the product is converted to the sodium salt and the excess salt removed by gel filtration.

Storage: -20°C


Ovalbumin

See Albumin, Egg, page 199

PABA

See p-Amino Benzoic Acid, page 199

Pancreatin 1X

Source: Porcine PancreasForm: White to off-white powder stabilized with

lactose.Product specifications:Protease: 1.0 - 2.0 x USPAmylase: 1.0 - 2.5 x USPLipase: 2.0 - 10.0 x USP units/mgLoss on Drying: 0.0 - 5.0%Fat: 0.0 - 3.0%Salmonella: NegativeE. coli: NegativeUnit Definition: Pancreatin will convert at least 25

times its weight of potato starch into soluble car-bohydrates in 5 minutes at 40°C and at least 25 times its weight of casein into proteases in

1 hour at 40°C.

19880 200 gm

Paraformaldehyde Solution, 4% in PBS

Description: Paraformaldehyde solution 4% in phos-phate buffered saline

Product specifications:Form: Colorless, clear solutionPurity (Formaldehyde): 91 - 93%pH (25°C): 7.0 - 7.6Storage: 4°C

19943 1 L

PBS, 10X Solution, pH 7.4ULTRAPURE

MB GradeForm: 10X PBS (Phosphate Buffered Saline) solution

consisting of 80.6 mM sodium phosphate, 19.4 mM potassium phosphate, 27 mM KCl, and 1.37 M NaCl in high purity dH2O, pH 7.4. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout. Dilute to 1X with high purity dH2O for a solution consisting of 10 mM phosphate, pH 7.4 ± 1 2.7 mM potassium chloride and 137 mM NaCl.

pH (25°C, 10X): 7.4 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes test

75889 100 ml 1 L 5 L




Polymerase (PN 71160).

77212 500 µl 2 x 500 µl





77119 500 µl

PCR Nucleotide Mix with dUTP, 10 mM SolutionULTRAPURE

Form: Premixed dNTPs at 10 mM each dATP, dCTP, dGTP, dUTP



77330 500 µl

PEG

See Polyethylene Glycol 400, page 235See Polyethylene Glycol 6000, page 235See Polyethylene Glycol 8000, page 235

Penicillin G, Potassium

USBioAnalyzedC16H17KN2O4SFormula Weight: 372.48Product specifications:Form: White powderAssay: 1440 - 1680 Penicillin G units/mg (as is)Identity A: By IRIdentity B: Passes testpH (60 mg/ml): 5.0 - 7.5Loss on Drying: ≤ 1.5%Crystallinity: Passes testMode of Action:

Interferes with bacterial cell wall synthesis and cell division.Inactivated by oxidizing and reducing agents, glycerols and alcohols.

CAS #113-98-4

19985 1 kg

Pepsin

(E.C.3.4.23.1)Source: Porcine StomachProduct specifications:Form: Lyophilized, salt free, 2X crystallizedActivity: ≥ 2500 units/mgUnit Definition: One unit causes an increase in

absorbance at 280 nm of 0.001/minute at 37°C, pH 2.0.

Storage: -20°CCAS #9001-75-6

20010 1 gm 5 gm 10 gm 25 gm

Biochemicals


Pepsin 1:10,000

(E.C.3.4.23.1)Source: Porcine StomachProduct specifications:Form: Cream colored powderActivity: 1:10,000 - 11,500Loss on Drying: ≤ 4.0%Storage: Protect from moisture.

20015 100 gm 500 gm 1 kg

Pepstatin A(Isovaleryl-Val-Val-4-Amino-3-Hydroxy-6-Methylheptanoyl-Ala-4-Amino-3-Hydroxy-6-Methylheptanoic Acid)

ULTRAPURE

C34H63N5O9

Formula Weight: 685.89Product specifications:Form: White powderChromatography (TLC): HomogeneousPeptide Content: ≥ 98%Biological Activity: ≥ 850 µg/mgApplication:� Pepstatin A, a protease inhibitor, inhibits Pepsin,

Renin, and Cathepsin D. Effective concentrations are 10-6 to 10-9 M. It does not inhibit Trypsin, Chymotrypsin, Papain, or Plasmin.

Note: Dissolves in methanol.Storage: 4°CCAS #26305-03-3

20037 5 mg 25 mg

Peptones

See Bacteriological Peptone, page 202See Casein Hydrolysate, page 205See Yeast Extract and Hydrolysate, pages 251

Phenol(Carbolic Acid)

ULTRAPURE

MB GradeC6H5OHFormula Weight: 94.11Product specifications:Form: Translucent crystalline solidAssay: ≥ 99%Clarity of Solution (5%, H2O): Passes testpH (5%, H2O): 4.5 - 6.0Freezing Point: ≥ 40.5°CWater: ≤ 0.1%Res. on Evaporation: ≤ 0.05%Buffer Equilibration Suitability: Passes testIron (Fe): ≤ 1 ppmLead (Pb): ≤ 2 ppmMagnesium (Mg): ≤ 1 ppmDNase (endo): Passes testPreservatives/Inhibitors: Passes testApplication:� Used primarily for DNA/RNA extraction and puri-

fication.Preparation of buffered phenol:1. Allow phenol to warm to room temperature and

then melt at 68°C.2. Add an equal volume of buffer (0.5 M Tris-HCl,

pH 8.0).3. Stir for about 15 minutes on a magnetic stirrer.4. When the two phases have separated, aspirate

the top (aqueous) phase.5. Add an equal volume of 0.1 M Tris-HCl, pH 8.0

and repeat stirring.6. Remove the aqueous phase and repeat extrac-

tions until the pH of the phenol is 7.8. (Note: Measure with pH paper).

7. After the phenol is equilibrated, remove the final aqueous phase. Top with 0.1 volumes of 0.1 M Tris-HCl, pH 8.0.

8. Store in a light-tight bottle at 4°C.Note: For convenience, we provide phenol which is

already equilibrated (PN 75829).Storage: -20°CPacked under inert atmosphereCAS #108-95-2

75830 500 gm

Phenol, pH 8.0, EquilibratedULTRAPURE

MB GradeForm: This product is supplied as water-saturated

phenol with a separate bottle of Phenol Equil-ibration Buffer. At the time of product use, simply add the entire contents of Phenol Equilibration Buffer to the water-saturated phenol.

pH (5%, after equilibration, 20°C): ≥ 8.0Contaminant Testing:

DNase: Passes testStorage: 4°CApplication: � Used in nucleic acid purification procedures and

dissolution of protein

75829 100 ml 400 ml

Phenol: Chloroform: Isoamyl Alcohol (25:24:1)ULTRAPURE

MB GradeForm: A liquid mixture of phenol, ACS Reagent

Grade chloroform and ACS Reagent Grade iso-amyl alcohol in a ratio of 25:24:1 supplied with an additional Alkaline Equilibration Buffer used to adjust the pH to 8.0.

Note: Phenol:Chloroform:Isoamyl Alcohol (25:24:1) is packaged at pH 6.7. The product is accompa-nied by a separate Alkaline Equilibration Buffer. When ready to use add the supplemental buffer’s entire contents to the Phenol:Chloroform:Isoamyl Alcohol, then mix or shake, and allow to separate. Once this is completed the product will be at a pH of 8.0.

Contaminant Testing: DNase: Passes test

pH (5%, H2O): 8.0Application: � Nucleic acid extraction and purificationPhenol/Chloroform RNA Isolation MethodA. Guanidine HCl Homogenization Buffer for RNA

Isolation: 8.0 M Guanidine HCl, 0.1 M Sodium Acetate, pH 5.5, 5.0 mM Dithiothreitol, 0.5% Sodium Lauryl Sarcosinate.

B. Extraction with Phenol: Most extractions are per-formed using equal volumes of equilibrated phe-nol with respect to the sample volume. Normally, the extraction is performed twice with phenol and once with a 1:1 solution of phenol/chloro-form. Hence, it is estimated that 25 ml of phenol is required for every 10 ml of sample.

Note: If regular phenol is purchased, it will need to be equilibrated before use. For this reason, many customers may prefer to purchase phenol that is already equilibrated (PN 75829).

C. RNA Resuspension Buffer: TE, pH 7.5.Storage: 4°C

75831 100 ml 400 ml

Biochemicals


Phenolphthalein

ACS Reagent GradeC20H14O4

Formula Weight: 318.32Product specifications:Form: White to off-white powderClarity of Alcohol Solution (1%, EtOH): Passes testVisual Transition:

From pH 8.0: Colorless topH 10.0: Red

CAS #77-09-8

20075 500 gm 1 kg

Phenol Red, Sodium Salt(Phenolsulfonphthalein)

C19H13O5SNaFormula Weight: 376.36Product specifications:Form: Rust/red crystalline powderIdentity: By IRClarity of Solution: Passes testVisual Transition Interval: pH 6.8 - Yellow pH 8.2 - RedCAS #34487-61-1

20085 10 gm 25 gm 50 gm

Phenolsulfonphthalein

See Phenol Red, page 234

DL-Phenylalanine

C6H5CH2CH(NH2)COOHFormula Weight: 165.19Product specifications:Form: White crystalline powderAssay: ≥ 99.0%Chromatagraphy (TLC): HomogeneousIdentity: By IRLoss on Drying: ≤ 0.3%Residue on Ignition: ≤ 0.3%Heavy Metals (Pb): ≤ 10 ppmAmmonia (NH3): ≤ 0.03%Arsenic (As): ≤ 3 ppmChloride (Cl): ≤ 0.02%Iron (Fe): ≤ 50 ppmCAS #150-30-1

20100 100 gm 1 kg

L-Phenylalanine

USBioAnalyzed(C6H5)CH2CH(NH2)COOHFormula Weight: 165.19Product specifications:Form: White crystalline powderAssay: 98.5 - 101.5% (dry basis)Chromatographic Purity: Passes testIdentity: By IRLoss on Drying: ≤ 0.3%[α]25

D : -32.7° to -34.7° (c=2 in H2O)pH (1%, H2O): 5.4 - 6.0Residue on Ignition: ≤ 0.4%Sulfate (SO4): ≤ 0.03%Heavy Metals (Pb): ≤ 0.0015%Chloride (Cl): ≤ 0.05%Iron (Fe): ≤ 0.003%Residual Solvents: Meets requirementsCAS #63-91-2

20105 100 gm 250 gm 1 kg 5 kg

Phenylmethyl Sulfonyl Fluoride(PMSF)

ULTRAPURE

C7H7FO2SFormula Weight: 174.19Product specifications:Form: Off-white/clear crystalsAssay: ≥ 99.0%Identity: By IRApplication: � PMSF is a protease inhibitor active against trypsin

and chymotrypsin.CAS #329-98-6

20203 5 gm 25 gm 100 gm

Phosphocreatine

See Creatine Phosphate, page 208

PIPES(Piperazine-N,N'-bis-[2-Ethanesulfonic Acid])

C8H18N2O6S2

Formula Weight: 302.37Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (20°C): 6.80 ± 0.15A (250 nm, 5% in 1N NaOH, 1 cm): ≤ 0.20Loss on Drying: ≤ 0.5%Iron (Fe): ≤ 30 ppmHeavy Metals (Pb): ≤ 10 ppmCAS #5625-37-6

20426 100 gm 1 kg

PIPES, Disodium Salt

C8H16N2O6S2Na2

Formula Weight: 346.33Product specifications:Form: White powderAssay: ≥ 98.0% (dry basis)Identity: By IRpKa (20°C): 6.80 ± 0.15A (250 nm, 0.1 M, 1 cm): ≤ 0.1Loss on Drying: ≤ 10.0%CAS #76836-02-7

20428 10 gm 100 gm 1 kg

PMSF

See Phenylmethyl Sulfonyl Fluoride, page 234

Polyanetholesulfonic Acid, Sodium Salt

(C10H11NaO4S)n

Formula Weight: (250.25)n

CAS #55963-78-5

20450 5 gm 100 gm

Biochemicals


Poly(dI-dC) • Poly(dI-dC) Sodium Salt

Product specifications:Description: Lyophilized, sodium salt complex of two

strands, each with an alternating sequence of deoxyinosinic acid and deoxycytidylic acid.

Form: White to off-white lyophilized powderConcentration: One A260 unit of double-stranded

polymers is ~ 50 µg.UV Absorption Spectrum: λmax: 248-252 nmAbsorbance ratios: A250/A260: 1.15-1.31 A280/A260: 0.64-0.74All UV Absorbance data were obtained in 0.1 M

sodium chloride, 0.02 M potassium phosphate, pH 7.0.

Note: To maintain a double-stranded configura-tion of the polymer in solution, rehydrate with a neutral buffer containing 100 mM NaCl. After reconstitution store the solution at -20°C.

Storage:-20°C


Polyethylene Glycol 400(PEG 400)

Formula Weight: 380 - 420Product specifications:Form: Clear colorless viscous liquidDensity (d20

4 ): 1.12 - 1.13Refractive Index (n20

d ): 1.465 - 1.469Viscosity (20°C): 102 - 120 mpasCAS #25322-68-3

19957 500 ml 1 L

Visit our website, usb.affymetrix.com, for the latest product information.

Polyethylene Glycol 6000, Flakes(PEG 6000)

Formula Weight: 5400 - 6600Product specifications:Form: White or slightly yellowish flakesSolubility (50%, Hot H2O): Clear and colorlessMelting Point: Approx. 60°CApplications:� Differential precipitation of DNA� Enhances hybridization rate of nucleic acids.� Improves efficiency of end-labeling with T4

Polynucleotide Kinase(1,2).� Used in ligation of blunt ended DNA.� Precipitates bacteriophage ± particles and plas-

mid DNA.� Used in preparing single-stranded M13 DNA.� Promotes cell fusion.PEG, with magnesium, causes DNA to undergo a

“psi” transition and it collapses into a highly condensed state(1). This results in macromolecular crowding(2) and increased efficiency in end-labeling.

References:1. Lerman, L. S. (1971) Proc Natl. Acad Sci, 68, 1886.2. Harrison, B. and Zimmerman, S. B., (1986) Anal

Biochem, 158, 307.CAS #25322-68-3

19972 1 kg 5 kg

Polyethylene Glycol 8000, Flakes(PEG 8000)

ULTRAPURE

Approx. M.W.=8,000Product specifications:Form: White flakesSolubility: Clear and colorlessMelting Point: 60 - 64°CApplications:� Differential precipitation of DNA� Enhances hybridization rate of nucleic acids.� Improves efficiency of end-labeling with T4



“psi” transition and it collapses into a highly condensed state(1). This results in macromolecular crowding(2) and increased efficiency in end-labeling and blunt ligation.


Biochem, 158, 307-315.CAS #25322-68-3

19959 500 gm 1 kg 5 kg

Polyethylene Glycol 8000, Powder(PEG 8000)

Molecular Weight Range: 7,000 - 9,000Product specifications:Form: White to off-white powderSolubility (50%, hot H2O): Clear and colorlessMelting Point: 60 - 64°CApplications:� Differential precipitation of DNA� Enhances hybridization rate of nucleic acids.� Improves efficiency of end-labeling with T4



“psi” transition and it collapses into a highly condensed state(1). This results in macromolecular crowding(2) and increased efficiency in end-labeling and blunt ligation.


Biochem, 158, 307.CAS #25322-68-3

19966 500 gm 1 kg 5 kg

Polymixin B Sulfate

USBioAnalyzedProduct specifications:Form: White to buff colored powderActivity:

≥ 6000 Polymixin B units/mg (dry basis)≥ 5580 Polymixin B units/mg (as is)

Identity (A, B, C): Passes testspH (5 mg/ml): 5.0 - 7.5Loss on Drying: ≤ 7.0%Content of Phenylalanine: 9%-12%CAS #1405-20-5

20551 10 mu 100 mu

Polyvinyl Pyrrolidone (PVP)

See PVP-K-30, page 237

Biochemicals


Ponceau S, Sodium SaltULTRAPURE

(C.I.27195)C22H12N4O13S4Na4

Formula Weight: 760.57Application: � As a protein stain for serum proteins separated by

agarose gel and cellulose acetate electrophoresisCAS #6226-79-5

32819 50 gm

Potassium ChlorideULTRAPURE

ACS Reagent GradeKClFormula Weight: 74.55Product specifications:Form: White crystalline powderAssay: 99.0 - 100.5%pH (5%, H2O, 25°C): 5.4 - 8.6Insoluble Matter: ≤ 0.005%Iodide (I): ≤ 0.002%Bromide (Br): ≤ 0.01%Chlorate & Nitrate (NO3): ≤ 0.003%Nitrogen Cmpds. (N): ≤ 0.001%Phosphate (PO4): ≤ 5 ppmSulfate (SO4): ≤ 0.001%Barium (Ba):Passes testHeavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 3 ppmCalcium (Ca): ≤ 0.002%Magnesium (Mg): ≤ 0.001%Sodium (Na): ≤ 0.005%CAS #7447-40-7

20598 500 gm 1 kg 5 kg

Potassium Chloride (KCl), 2 M SolutionULTRAPURE

MB GradeForm: 2 M solution of potassium chloride (KCl) in

high purity dH2O. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles.

Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testCAS #7447-40-7

75896 100 ml

Potassium Phosphate, DibasicULTRAPURE

ACS Reagent GradeK2HPO4

Formula Weight: 174.18Product specifications:Form: White crystalline powderAssay: ≥ 98.5%pH (5%, H2O, 25°C): 8.5 - 9.6Insoluble Matter: ≤ 0.01%Loss on Drying: ≤ 1.0%Chloride (Cl): ≤ 0.003%Nitrogen Compds. (N): ≤ 0.001%Sulfate (SO4): ≤ 0.005%Heavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 0.001%Sodium (Na): ≤ 0.05%Storage: Protect from moisture (product is deliques-

cent).CAS #7758-11-4

20274 500 gm 1 kg 2.5 kg

Potassium Phosphate, Dibasic(Dipotassium Phosphate)

CP GradeK2HPO4

Formula Weight: 174.18Product specifications:Form: White deliquescent powderAssay: ≥ 98.0% K2HPO4 (dry basis)Identity: By IRArsenic (As): ≤ 3 ppmFluoride (F): ≤ 10 ppmInsol. Substances: ≤ 0.2%Lead (Pb): ≤ 2 ppmLoss on Drying: ≤ 2.0%Note: This product is a general laboratory reagent

suitable for most laboratory animal diets where trace element deficiencies are not required. One molar stock solutions of this product may be slightly hazy. Where use of a high quality research buffer is required, we recommend the use of our ACS Reagent Grade phosphate buffers.

Storage: Dessicate (product is deliquescent).CAS #7758-11-4

20276 1 kg 10 kg

Potassium Phosphate, Monobasic, AnhydrousULTRAPURE

ACS Reagent GradeKH2PO4

Formula Weight: 136.09Product specifications:Form: White crystalline powderAssay: ≥ 99.0%pH (5%, H2O, 25°C): 4.1 - 4.5Insoluble Matter: ≤ 0.01%Loss on Drying: ≤ 0.2%Chloride (Cl): ≤ 0.001%Nitrogen Compds. (N): ≤ 0.001%Sulfate (SO4): ≤ 0.003%Heavy Metals (Pb): ≤ 0.001%Iron (Fe): ≤ 0.002%Sodium (Na): ≤ 0.005%CAS #7778-77-0

20227 500 gm 1 kg

Potassium Phosphate, Monobasic

KH2PO4

Formula Weight: 136.09Product specifications:Form: White crystalline powderAssay: ≥ 98.0% (dry basis)Arsenic (As): ≤ 3 ppmFluoride (F): ≤ 10 ppmInsoluble Substances: ≤ 0.2%Lead (Pb): ≤ 2 ppmLoss on Drying: ≤ 1%Note: This product is a general laboratory reagent


CAS #7778-77-0

20228 1 kg 10 kg

Biochemicals


Protamine Sulfate

Source: SalmonProduct specifications:Form: White to off-white powderAssay: Each mg neutralizes not less than 100 USP

Heparin Units (dry basis)Loss on Drying: ≤ 5.0%Nitrogen (N): 22.5 - 25.5% (dry basis)Sulfate (SO4): 16.0 - 22.0% (dry basis)CAS #9009-65-8

20795 25 gm 100 gm

Proteinase K

Source: Tritirachium albumProduct specifications:Form: Lyophilized powderConcentration: 20 - 50 units/mgMolecular Weight: 28.9 kDaUnit Definition: One unit is the amount of enzyme

that liberates folin positive amino acids and peptides corresponding to 1 µmol tyrosine in 1 minute at 37°C using hemoglobin as substrate.

Application: � Proteinase K inactivates DNase and RNase from

most mammalian cells and microorganisms in the presence of 0.5 - 1% SDS. It can also be used for research involving protein structure because of its cleavage specificity.

Storage: Shipped on cold pack. Store dry at 4°C.CAS #39450-01-6

76230Y 100 mg76230Z 500 mg76230 1 gm (2 x 500 mg)

pUC 19 DNA

See Molecular Biology Products & Kits section, page 129

Purification Solutions

Phenol, pH 8.0, Equilibrated, page 233Phenol:Chloroform:Isoamyl Alcohol (25:24:1),

page 233

PVP-K-30(Polyvinylpyrrolidone)

Formula Weight: 40,000 (average)Product specifications:Form: Off-white powderActive content: ≥ 95%K-value (viscosity of 1% solids w/v aqueous solu-

tion): 26 - 34Moisture: ≤ 5.0%% Unsaturation (calculated as % vinylpyrrolidone):

≤ 0.5%CAS #9003-39-8

20611 1 kg

Pyruvic Acid, Sodium Salt(Sodium Pyruvate)

CH3COCOONaFormula Weight: 110.04Product specifications:Form: White to slightly yellow crystalline powderAssay: ≥ 98.5%Identity: By IRMoisture (KF): ≤ 1.0%Chloride (Cl): ≤ 30 ppmSulfate (SO4): ≤ 0.01%Heavy Metals (Pb): ≤ 10 ppmCAS #113-24-6

20995 10 gm 25 gm 100 gm 500 gm

D-(+)-Raffinose, Pentahydrate

C18H32O16•5H2O


D : +104.9° ± 1.3° (c=4 in H2O & NH3) (cor-rected to a theoretical pentahydrate)

Loss on Drying: ≤ 15.3%Residue on Ignition: ≤ 0.05%Iron (Fe): ≤ 5 ppmHeavy Metals (Cu): ≤ 10 ppmArsenic (As): ≤ 0.5 ppmCAS #17629-30-0

21060 100 gm 500 gm 1 kg

RapidGel 40% Liquid Acrylamide Stock SolutionULTRAPURE

MB GradeForm: Pre-mixed, 40% liquid acrylamide (19:1

acrylamide:bis-acrylamide ratio) in ready-to-use form for Polyacrylamide Gel Electrophoresis (PAGE). Stock solution provides flexibility for dif-ferent acrylamide percentages in a wide variety of gel applications. This formulation does not contain urea.

Testing: Functionally tested in gel electrophoresis.To use: Dilute 40% stock solution to desired con-

centration with appropriate running buffer for PAGE. To make 100 ml of a sequencing gel solu-tion containing 7-8.3 M urea, add an appropriate volume of 40% RapidGel solution, 10 ml of 10X (or 5 ml of 20X) running buffer, 42-50 gm of urea, and high purity dH2O to 100 ml in a clean, graduated cylinder. Stir and warm solution at 35-45°C to dissolve urea. (Do not microwave.) Cool to room temperature. Filter through a 0.2 µm filter and leave under gentle vacuum for 10 minutes to de-gas. Add 600 µl 10% ammoni-um persulfate and 60 µl TEMED per 100 ml gel solution to polymerize. Allow to polymerize at room temperature from one hour to overnight. For polymerization within 30-45 minutes, add 1 ml 10% ammonium persulfate and 100 µl TEMED per 100 ml gel solution. Pour gel immediately.

Storage: 4°C

75848 500 ml

Biochemicals


RapidGel-XL 6% Liquid Acrylamide DNA Sequencing Gel, TBE FormulationULTRAPURE

MB GradeForm: Pre-mixed, liquid acrylamide in ready-to-

use form for Ultrapure DNA sequencing gels. Contains TBE Buffer and 7 M urea. The most commonly requested gel formulation, RapidGel-XL, 6%, is a high resolution matrix for DNA sequenc-ing, yielding up to 40% more readable sequence per gel when using either radioactive or fluores-cent labeling methods. No gel fixing is necessary.

Testing: Functionally tested in sequencing gel elec-trophoresis.

To use: Intended for use with 1X TBE Buffer. De-gas solution by placing under gentle vacuum for 10 minutes at room temperature. Add 600 µl 10% ammonium persulfate and 60 µl TEMED per 100 ml gel solution to polymerize. Allow to polymerize at room temperature from one hour to overnight. For polymerization within 30-45 minutes, add 1 ml 10% ammonium persul-fate and 100 µl TEMED per 100 ml gel solution. Pour gel immediately. Do not fix gel.

Note: Urea may crystallize out of solution when the gel mix is stored as recommended at 4°C. This is normal and will not affect performance. If this occurs, warm the gel mix at 37°C for 10 minutes or until the urea redissolves. Mix gently. Cool to room temperature before adding the ammonium persulfate and TEMED.

Use with corresponding TBE Running Buffer: 10X Ready Mixed Powder, PN 70454, packaged in 6 x 200 ml bottles or TBE Buffer, 5X Solution, PN 75891, packaged in 1 and 5 liter sizes.

Storage: 4°C

75861 500 ml

RapidRun Agarose Buffer, 20X SolutionULTRAPURE

MB GradeForm: RapidRun Agarose Buffer allows for gels to be

run in one-fourth the time required for standard buffers, such as TAE and TBE. The buffer is a non-borate weak acid that enables high voltage and more rapid electrophoretic runs. Temperature seldom exceeds 38°C during high voltage runs, minimizing the band diffusion and gel degrada-tion commonly associated with TAE and TBE under the same high voltage conditions.

20X RapidRun Agarose Buffer consists of 1.78 M Tris, 0.57 M Taurine, and 0.01 M EDTA. The solu-tion is 0.2 µm filtered.

pH (1:20, 25°C): 8.9 + 0.1Conductivity (1X): 830 + 100 µmhos/cmTesting: Meets or exceeds functional test criteria for

high voltage (300v, 15 minutes) mini-agarose gel electrophoresis.

Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test

Applications:� High voltage, fast agarose gel electrophoresis

(20-30 volts/cm).� This buffer is compatible with commercial gel

extraction kits. However, the pH should be adjust-ed with sodium acetate, pH 5.0, as recommended by the manufacturer.

Note: 1 ml of RapidRun Loading Dye (PN 77524) is included with each pack size. The dye front migrates lower than 50 bases.

Storage: Ambient. Crystals which form at low tem-peratures can be re-dissolved by warming briefly.

77523 1 L 5 L

RapidRun Loading DyeULTRAPURE

MB GradeForm: 5X loading dye (0.05% dye in 50% glycerol).Contaminant Testing:

DNase (endo): Passes testDNase (exo): Passes testRNase: Passes test

Application: � Used as a loading dye designed for high voltage

agarose gel electrophoresis with buffers such as RapidRun Agarose Buffer (PN 77523). It may also be used for standard voltage gel electro-phoresis. RapidRun Loading Dye develops an intense orange color in the presence of RapidRun Agarose Buffer. The dye front migrates lower than 50 bases.

77524 1 ml 5 ml

Riboflavin(Vitamin B2 ; Vitamin G)


Formula Weight: 376.36Product specifications:Form: Yellow-orange crystalline powderAssay: 98.0 - 102.0% (dry basis)Identity: By IR[α]25

D : -115° to -135° (c=0.5 in 0.05 M NaOH)Loss on Drying: ≤ 1.5%Residue on Ignition: ≤ 0.3%Limit of Lumiflavin: Passes testResidual Solvents: Meets requirementsStorage: Protect from light.CAS #83-88-5

21145 25 gm 100 gm 500 gm 1 kg

D-(–)-Ribose

C5H10O5

Formula Weight: 150.13Product specifications:Form: White powderAssay: ≥ 98.0% (dry basis)[α]20

D : -20.4° ± 0.4° (c=2 in H2O, NH3)Loss on Drying: ≤ 0.5%Residue on Ignition: ≤ 0.1%Arsenic (As): ≤ 0.5 ppmIron (Fe): ≤ 5 ppmHeavy Metals (Cu): ≤ 10 ppmCAS #50-69-1

21220 10 gm 100 gm

Rifampin

C43H58N4O12

Formula Weight: 822.94Product specifications:Form: Reddish brown crystalline powderAssay:

95.0 - 103.0% of C43H58N4O12 (dry basis) 93.0 - 103.0% of C43H58N4O12 (as is)Identity: By IRpH (1%, H2O): 4.5 - 6.5Loss on Drying: ≤ 2.0%Related Substances: Passes testMode of Action: Inhibits RNA synthesis by binding

to and inhibiting the β subunit of RNA polym-erase.

Storage: Light-resistant containers, protect from heat.

CAS #13292-46-1

21246 1 gm 5 gm

RNA, Free Acid

Source: Torula YeastProduct specifications:Form: White powderMoisture: ≤ 8.0%Nitrogen: ≥ 15.3% (dry basis)Phosphorous: ≥ 9.0% (dry basis)CAS #73049-77-1

21185 100 gm 1 kg

Biochemicals


RNA, Sodium Salt

Source: YeastProduct specifications:Form: White powderMoisture: 4.0 - 8.0%Phosphorous: ≥ 8.0%Nitrogen: ≥ 14.5%pH (1% H2O): 6.5 ± 0.5Biuret test: NegativeCAS #73049-77-1

21190 25 gm 100 gm 1 kg

RNase A, Lyophilized

(E.C. 3.1.4.22)Source: Bovine pancreasForm: Lyophilized powderUnit Definition:One unit is that amount of enzyme

causing the hydrolysis of RNA at a rate such that K equals unity at 25°C and pH 5.0.

Storage: -20°C, desiccatedCAS #9001-99-4

21195 100 mg 1 gm 5 gm

SAMe-PTS

See S-Adenosyl-L-Methionine Sulfate p-Toluenesulfonate, page 190

SDS


DL-Serine (Non-Animal)

CH2OHCHNH2COOHFormula Weight: 105.09Product specifications:Form: White crystalline powderAssay: ≥ 98.5% (dry basis)Solubility (2%, H2O): Clear, colorless solutionIdentity: By IRLoss On Drying: ≤ 0.3%Residue on Ignition: ≤ 0.1%Chloride (Cl): ≤ 0.021%Sulfate (SO4): ≤ 0.05%Heavy Metals (Pb): ≤ 20 ppm

21486 100 gm 1 kg

Sodium Acetate (NaOAc), 3 M Solution, pH 5.5ULTRAPURE

MB GradeForm: 3 M solution of sodium acetate (NaOAc) in

high purity dH2O, pH 5.5. Solution is 0.2 µm fil-tered and dispensed into durable, square Nalgene bottles.

pH: 5.5 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test

75897 100 ml 500 ml

Sodium Aspartate

See Aspartic Acid, Sodium Salt, page 201

Sodium AzideULTRAPURE

NaN3

Formula Weight: 65.01Product specifications:Form: White crystalline powderAssay: ≥ 99.0%Loss on Drying: ≤ 0.5%CAS #26628-22-8

21610 100 gm 500 gm

Sodium Cacodylate

See Cacodylic Acid, Sodium Salt, page 204

Sodium Carbonate, Anhydrous

ACS Reagent GradeNa2CO3

Formula Weight: 105.99Product specifications:Form: White crystalline powderAssay: ≥ 99.5%Insoluble Matter: ≤ 0.01%Loss on Drying (285°C): ≤ 1.0%Chloride (Cl): ≤ 0.001%Nitrogen Compounds (N): ≤ 0.001%Phosphate (PO4): ≤ 0.001%Silica (SiO4): ≤ 0.005%Sulfur Compounds (SO4): ≤ 0.003%Ammonium Hydroxide Precipitate: ≤ 0.01%Heavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 5 ppmCalcium (Ca): ≤ 0.03%Magnesium (Mg): ≤ 0.005%Potassium (K): ≤ 0.005%CAS #497-19-8

21616 500 gm 2.5 kg

Sodium Caseinate

See Casein, Sodium, page 205

Sodium Chloride (NaCl), 5 M SolutionULTRAPURE

MB GradeForm: 5 M solution of sodium chloride (NaCl) in high

purity dH2O. Solution is 0.2 µm filtered and dis-pensed into durable, square Nalgene bottles.

Contaminant Testing: Heavy Metals: ≤ 5 ppm DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes test

75888 100 ml 500 ml 1 L

Sodium ChlorideULTRAPURE

ACS Reagent GradeNaClFormula Weight: 58.44Product specifications:Form: White/clear crystalline powderAssay: ≥ 99.0%pH (5%, H2O, 25°C): 5.0 - 9.0Insoluble Matter: ≤ 0.005%Iodide (I): ≤ 0.002%Bromide (Br): ≤ 0.01%Chlorate & Nitrate (NO3): ≤ 0.003%Nitrogen Cmpds. (N): ≤ 0.001%Phosphate (PO4): ≤ 5 ppmSulfate (SO4): ≤ 0.004%Barium (Ba): Passes testHeavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 2 ppmCalcium (Ca): ≤ 0.002%Magnesium (Mg): ≤ 0.001%Potassium (K): ≤ 0.005%Application: � Used for the precipitation of DNA.CAS #7647-14-5

21618 500 gm 1 kg 5 kg 10 kg

Sodium Citrate

See Citric Acid, Trisodium Salt, page 207

Sodium Deoxycholate

See Deoxycholic Acid, Sodium Salt, page 210

Biochemicals


Sodium Dodecyl Sulfate (SDS)(Sodium Lauryl Sulfate)

ULTRAPURE

CH3(CH2)11OSO3NaFormula Weight: 288.38Product specifications:Form: White crystalline flakesAssay: ≥ 99% (acidity)Assay: ≥ 98% (GC)A (280 nm, 3%, 1 cm): ≤ 0.1A (230 nm, 3%, 1 cm): ≤ 0.2Loss on Drying: ≤ 1%Insoluble Matter: ≤ 0.003%Chloride (Cl): ≤ 0.01%Phosphate (PO4): ≤ 1 ppmCopper (Cu): ≤ 5 ppmIron (Fe): ≤ 1 ppmLead (Pb): ≤ 5 ppmProtocol for 10% SDS Solution: SDS: 100 gm H2O: 900 ml Heat to 68° to dissolve. Adjust to pH 7.2 and

bring to 1 liter. There is no need to sterilize 10% SDS.

CAS #151-21-3

75819 100 gm 1 kg

Sodium Dodecyl Sulfate (SDS)(Sodium Lauryl Sulfate)

CH3(CH2)11OSO3NaFormula Weight: 288.38Product specifications:Form: White crystalline powderIdentity: By IRAssay: ≥ 95% (dry basis)Solubility (20%, H2O): Clear, faintly yellow solutionCAS #151-21-3

18220 500 gm 1 kg 5 kg

Sodium Dodecyl Sulfate (SDS), 10% SolutionULTRAPURE

Form: 10% (w/v) solution of sodium dodecyl sulfate (SDS) in high purity dH2O. Solution is 0.2 µm fil-tered and dispensed into durable, square Nalgene bottles.

A (280 nm, 3%, 1 cm): ≤ 0.1A (230 nm, 3%, 1 cm): ≤ 0.2Note: Precipitation of SDS is not unusual. Warm

gently to redissolve.

77504 100 ml 500 ml 1 L

Sodium Dodecyl Sulfate (SDS), 20% SolutionULTRAPURE

Form: 20% (w/v) solution of sodium dodecyl sulfate (SDS) in high purity dH2O. Solution is 0.2 µm fil-tered and dispensed into durable, square Nalgene bottles.

A (280 nm, 3%, 1 cm): ≤ 0.1A (230 nm, 3%, 1 cm): ≤ 0.2Note: Precipitation of SDS is not unusual. Warm

gently to redissolve.

75832 100 ml 500 ml

Sodium Glutamate

See Glutamic Acid, Monosodium Salt, page 218

Sodium-N-Lauryl Sarcosine

See N-Lauryl Sarcosine, Sodium Salt, page 224

Sodium Lauryl Sulfate


Sodium Phosphate, Dibasic Anhydrous

ACS Reagent GradeHygroscopicNa2HPO4

Formula Weight: 141.96Product specifications:Form: White crystalline powderAssay: ≥ 99.0%pH (5%, H2O, 25°C): 8.7 - 9.3Insoluble Matter: ≤ 0.01%Loss on Drying : ≤ 0.2%Chloride (Cl): ≤ 0.002%Sulfate (SO4): ≤ 0.005%Heavy Metals (Pb): ≤ 0.001%Iron (Fe): ≤ 0.002%Storage: Store ambient, protect from moisture

(product is hygroscopic).CAS #7558-79-4

21660 500 gm 2.5 kg

Sodium Phosphate, Dibasic, HeptahydrateULTRAPURE

ACS Reagent GradeEffloresces in warm dry airNa2HPO4

•7H2OFormula Weight: 268.07Product specifications:Form: White powderAssay: 98.0 - 102.0%pH (5%, H2O, 25°C): 8.7 - 9.3Insoluble Matter: ≤ 0.005%Chloride (Cl): ≤ 0.001%Nitrogen Compounds (N): ≤ 0.001%Sulfate (SO4): ≤ 0.005%Heavy Metals (Pb): ≤ 0.001%Arsenic (As): ≤ 5 ppmIron (Fe): ≤ 0.001%Note: While this product is a high quality, analytical

reagent grade buffer, as a good general labora-tory practice we recommend filtering solutions before use.

Storage: Store in tightly closed containers.CAS #7782-85-6

20232 500 gm 1 kg

Sodium Phosphate, Monobasic(Monosodium Phosphate)

CP GradeNaH2PO4

Formula Weight: 119.98Product specifications:Form: White, slightly hygroscopic crystalline powderAssay: 98.0 - 103.0% (NaH2PO4, dry basis)Identity: Passes testsArsenic (As): ≤ 3 ppmFluoride (F): ≤ 0.005%Insoluble Substances: ≤ 0.2%Lead (Pb): ≤ 4 ppmLoss on Drying: ≤ 2.0%Note: This product is a general laboratory reagent


Storage: Ambient, desiccate (product is slightly hygroscopic).

CAS #7558-80-7

20229 1 kg 5 kg

Biochemicals


Sodium Phosphate, Monobasic, MonohydrateULTRAPURE

ACS Reagent GradeSlightly HygroscopicNaH2PO4

•H2OFormula Weight: 137.99Product specifications:Form: White crystalline powderAssay: 98.0 - 102.0%pH (5%, H2O, 25°C): 4.1 - 4.5Insoluble Matter: ≤ 0.01%Chloride (Cl): ≤ 5 ppmSulfate (SO4): ≤ 0.003%Calcium (Ca): ≤ 0.005%Potassium (K): ≤ 0.01%Heavy Metals (Pb): ≤ 0.001%Iron (Fe): ≤ 0.001%CAS #10049-21-5

20233 500 gm 1 kg

Sodium Pyruvate

See Pyruvic Acid, Sodium Salt, page 237

Sodium Selenite

Na2SeO3

Formula Weight: 172.94Product specifications:Form: White to off-white powderStorage: Ambient; hygroscopic, store in tightly

closed containers in a cool, dry place. The com-pound is very readily reduced to red elemental selenium in the presence of small amounts of reducing agents, and should not be allowed to come into contact with organic material or even dust particles.

CAS #10102-18-8

21665 100 gm 500 gm

Sodium Succinate

See Succinic Acid, Disodium Salt, page 242

Soluble Starch

See Starch-Soluble, page 242

D-SorbitolULTRAPURE


Formula Weight: 182.17Product specifications:Form: White crystalline powderAssay (dry basis): 91.0 - 100.5%pH (10%, H2O): 3.5 - 7.0Identity: By IRIdentity (A, B): Passes testsMicrobial Limits: Passes testMoisture (KF): ≤ 1.5%Residue on Ignition: ≤ 0.1%Reducing Sugars: Passes testLimit of Nickel: Passes testStorage: In tightly closed containers, material is

hygroscopic.CAS #50-70-4

21710 500 gm 1 kg 5 kg

Spermidine, TrihydrochlorideULTRAPURE

C7H19N3•3HCl

Formula Weight: 254.63Product specifications:Form: White to pale pink crystalline powderAssay: ≥ 99%Identity: By IRApplications:� Causes a 2-fold increase in full length cDNA with

or without pyrophosphate or Actinomycin D using AMV Reverse Transcriptase.

� Used in cosmid cloning.� Often included in T4 Polynucleotide Kinase stocks.Storage: -20°CCAS #334-50-9

21760 1 gm 5 gm

Spermine, Tetrahydrochloride

C10H26N4•4HCl

Formula Weight: 348.18Product specifications:Form: White powderIdentity: By IRStorage: -20°CCAS #306-67-2

21755 5 gm

SSC, 20X SolutionULTRAPURE

Form: 20X SSC solution consisting of 0.3 M triso-dium citrate and 3.0 M sodium chloride in high purity dH2O, pH 7.0. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright car-ton with a convenient dispensing spout.

pH (25°C, 20X): 7.0 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testApplication: � Used in nucleic acid blotting and hybridization

techniques.

19629 500 ml 1 L 5 L

SSPE, 20X SolutionULTRAPURE

MB GradeForm: 20X SSPE solution consisting of 3 M NaCl,

200 mM sodium phosphate and 20 mM EDTA in high purity dH2O, pH 7.4. Solution is 0.2 µm fil-tered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout.

pH (25°C, 20X): 7.4 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testStorage: -20°C

75890 1 L 5 L

Biochemicals


Starch, HydrolyzedULTRAPURE

Source: PotatoProduct specifications:Form: White to off-white powderResidue on Ignition: ≤ 1%Loss on Drying: ≤ 20%Gel Strength: Passes testNote: Typically a 12-14% gel is recommended for

electrophoresis. Gel consistency is lot specific and depends on the percentage of amylose and amylopectin in starch, which varies naturally. Optimum concentration must be determined for each lot by the user.

CAS #9005-25-8

32823 100 gm 1 kg

Starch-SolubleULTRAPURE

ACS Reagent GradeSource: PotatoProduct specifications:Form: White powderSolubility: Passes testpH (2%, H2O, 25°C): 5.0 - 7.0Loss on Drying: ≤ 12%Sensitivity: Passes testResidue on Ignition: ≤ 0.4%CAS #9005-84-9

21695 1 kg

Stock Solutions

Ammonium Acetate (NH4OAc), 5 M Solution, page 199

Betaine, 5 M Solution, page 202Dithiothreitol (DTT) 0.1 M Solution, page 213EDTA, 0.5 M Solution, page 215Lithium Chloride (LiCl), 7.5 M Solution, page 225Magnesium Chloride (MgCl2), 1 M Solution,

page 226Potassium Chloride (KCl), 2 M Solution, page 236Sodium Acetate (NaOAc), 3 M Solution, pH 5.5,

page 239Sodium Chloride (NaCl), 5 M Solution, page 239Sodium Dodecyl Sulfate (SDS), 10% Solution,

page 240Sodium Dodecyl Sulfate (SDS), 20% Solution,

page 240Water, Nuclease-Free, page 250Water, RNase-Free, DEPC Treated, page 250

Stop SolutionULTRAPURE

Contents:95% Formamide20 mM EDTA0.05% Bromophenol Blue0.05% Xylene Cyanol FF

Application: � Used to stop and load sequencing reaction prod-

ucts onto denaturing acrylamide gels.Storage: -20°C

70724 1,200 µl

StreptavidinULTRAPURE

Source: E. coli recombinant proteinProduct specifications:Form: White, lyophilized powderActivity: ≥ 17 units/mgSolubility (0.1 M NaP, pH 7.4): ≥ 5 mg/mlSDS PAGE: Single bandUnit Definition: One unit is that amount of protein

required to bind one microgram of D-biotin.Applications:� A tetrameric protein containing four high affinity

binding sites for biotin.� Used with biotin in the purification of DNA-

binding proteins.Storage: 4°C, desiccated; -20°C, reconstituted.

11681 5 mg 10 mg

Streptavidin-Alkaline Phosphatase Conjugate

Storage: 4°C

11687 250 µl 1,500 µl

Streptomycin SulfateULTRAPURE

USBioAnalyzed(C21H39N7O12)2

•3H2SO4

Formula Weight: 1457.38Product specifications:Form: White to off-white powderActivity: 650 µg - 850 µg of C21H39N7O12/mg (as is)Identity (A, B): Passes testspH (200 mg streptomycin/ml): 4.5 - 7.0Loss on Drying: ≤ 5.0%Mode of Action: Inhibits protein synthesis by binding

to the S12 protein of the 30S ribosomal subunit.CAS #3810-74-0

21865 10 gm 100 gm 1 kg

Succinic Acid, Free Acid

HOOCCH2CH2COOHFormula Weight: 118.09Product specifications:Form: White crystalline powderAssay: ≥ 99.0%Identity: By IRMelting Point: 185 - 190°CMoisture (KF): ≤ 1.0%Iron (Fe): ≤ 5 ppmHeavy Metals: ≤ 10 ppmResidue on Ignition: ≤ 0.025%CAS #110-15-6

21900 500 gm 1 kg 5 kg

Succinic Acid, Disodium Salt, Hexahydrate(Sodium Succinate)

C4H4Na2O4•6H2O

Formula Weight: 270.14Product specifications:Form: White crystalline powderAssay: 97.0 - 102.0% (as is)Identity: By IRLoss on Drying: ≤ 40.0%Storage: Store in tightly closed containers.CAS #6106-21-4

21910 500 gm

Biochemicals


SucroseULTRAPURE

C12H22O11

Formula Weight: 342.30Product specifications:Form: White/clear crystalline powder[α]25

D : +66.3° to +66.8° (c=26 in H2O)A (260 nm, 1 M, 1 cm): ≤ 0.20Insoluble Matter: ≤ 0.005%Loss on Drying: ≤ 0.03%Residue on Ignition: ≤ 0.01%Titratable Acid: ≤ 0.0008 meq/gmInvert Sugar: ≤ 0.05%Chloride (Cl): ≤ 0.005%Sulfate and Sulfite (SO4): ≤ 0.005%Arsenic (As): ≤ 1 ppmBarium (Ba): ≤ 1 ppmCadmium (Cd): ≤ 1 ppmCalcium (Ca): ≤ 5 ppmCopper (Cu): ≤ 1 ppmIron (Fe): ≤ 1 ppmLead (Pb): ≤ 1 ppmMagnesium (Mg): ≤ 1 ppmManganese (Mn): ≤ 1 ppmZinc (Zn): ≤ 1 ppmContaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes testCAS #57-50-1

21938 1 kg 5 kg

Sucrose

ACS Reagent GradeC12H22O11

Formula Weight: 342.30Product specifications:Form: White/clear crystalline powder[α]25

D : +66.3° to +66.8° (c=26, H2O)Insoluble Matter: ≤ 0.005%Loss on Drying: ≤ 0.03%Residue on Ignition: ≤ 0.01%Titratable Acid ≤ 0.0008 meq/gmChloride (Cl): ≤ 0.005%Sulfate and Sulfite (SO4): ≤ 0.005%Heavy Metals (Pb): ≤ 5 ppmIron (Fe): ≤ 5 ppmInvert Sugar: ≤ 0.05%CAS #57-50-1

21931 500 gm 2.5 kg

Sucrose, Crystalline

C12H22O11

Formula Weight: 342.30Product specifications:Form: White/clear crystalsIdentity: By IR[α]20

D : +65.9° to +67.1° (c=26 in H2O)Solubility (26%, H2O): Clear and colorlessCAS #57-50-1

21930 1 kg 5 lb 100 lb

Sulfanilic Acid

NH2C6H4SO3HFormula Weight: 173.19Product specifications:Form: White to pale cream colored powderAssay: 98.0 - 102.0% (as is)Identity: By IRCAS #121-57-3

21985 500 gm

TAE Buffer, 10X SolutionULTRAPURE

Form: 10X TAE solution consisting of 400 mM Tris and 0.01 M EDTA in high purity dH2O adjusted to pH 8.3. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout. Dilute to 1X with high purity dH2O for a solution consisting of 40 mM Tris Acetate, pH 8.3 and 1.0 mM EDTA.

pH (25°C, 10X): 8.3 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test

75904 1 L 5 L

TAE Buffer, 50X SolutionULTRAPURE

Form: 50X TAE solution consisting of 2.0 M Tris and 0.05 M EDTA in high purity dH2O and adjusted to pH 8.3. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. Dilute to 1X with high purity dH2O for a solution consisting of 40 mM Tris Acetate, pH 8.3 and 1.0 mM EDTA.

pH (25°C, 50X): 8.3 ± 0.1

74015 100 ml

Taurine(2-Amino Ethane Sulfonic Acid)

ULTRAPURE

MB GradeC2H7NO3SFormula Weight: 125.15Product specifications:Form: White crystalline powderAssay: ≥ 98.5%Identity: By IRSolubility (0.5 M, H2O): Clear and colorlesspH (5%, H2O): 4.0 - 7.0A (260 nm, 0.5 M, 1 cm): ≤ 0.2A (280 nm, 0.5 M, 1 cm): ≤ 0.2Residue on Ignition: ≤ 0.1%Heavy Metals: ≤ 10 ppmAmmonia (NH3): ≤ 0.02%Arsenic (As): ≤ 2 ppmChloride (Cl): ≤ 0.01%Sulfate (SO4): ≤ 0.01%Use Test: Functionally tested for electrophoresis.CAS #107-35-7

75824 1 kg

TBE Buffer, 5X SolutionULTRAPURE

Form: 5X TBE (Tris-Borate-EDTA) solution consisting of 0.445 M Tris, 0.445 M boric acid and 0.01M EDTA. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout. Dilute to 1X with high purity water for a solution consisting of 0.089 M Tris, 0.089 M boric acid and 0.002 M EDTA.

Testing: Meets or exceeds functional test criteria for high quality gel electrophoresis.

Application: � Primarily used as a buffer in gel electrophoresis

procedures.

75891 1 L 5 L

TBE Buffer, 10X Ready-Mixed PowderULTRAPURE

Form: Pre-weighed, pre-mixed powdered buffer packaged in 200 ml bottles for reconstitution to a 10X solution. Reconstitute with high purity dH2O to 200 ml. After reconstitution, the 10X solution consists of 0.89 M Tris, 0.89 M boric acid and

0.02 M EDTA. Each pack contains 6 x 200 ml bottles. Total volume is 1.2 liters of 10X buffer or 12 liters of 1X buffer.

Testing: Meets or exceeds functional test criteria for high quality gel electrophoresis.

Application: � Primarily used as a buffer in gel electrophoresis

procedures.Storage: Ambient. The reconstituted buffer is stable

for several days at 4°C if 0.2 µm filtered.

70454 6 x 200 ml

Biochemicals


TBS, 20X Solution, pH 7.4ULTRAPURE

MB GradeForm: 20X TBS (Tris Buffered Saline) solution con-

sisting of 500 mM Tris, 60 mM KCl and 2.8 M NaCl in high purity dH2O, pH 7.4. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout. Dilute to 1X with high purity water for a solution consisting of 25 mM Tris, pH 7.4, 3.0 mM KCl, and 140 mM NaCl.

pH (25°C, 20X): 7.4 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testApplication: � Used in Western blotting protocols.

75892 100 ml 1 L 5 L

TBS with Tween (TBST), 20X Solution, pH 7.4ULTRAPURE

MB GradeForm: 20X TBS (Tris Buffered Saline) solution con-

sisting of 500 mM Tris, 60 mM KCl, 2.8 M NaCl, and 1.0% Tween 20 in high purity dH2O adjusted to pH 7.4. Solution is 0.2 µm filtered and is supplied in an upright carton with a convenient dispensing spout. Dilute to 1X with high purity water for a solution consisting of 25 mM Tris, pH 7.4, 3.0 mM KCl, 140 mM NaCl and 0.05% Tween 20.

pH (25°C, 20X): 7.4 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test Protease: Passes testApplication: � Used in Western blotting protocols.

77500 1 L 5 L

TCEP-HCI, 0.5 M Solution, pH 6.6 ± 0.1(Tris [Carboxyethyl] Phosphine Hydrochloride)

Form: TCEP-HCI solution consisting of 0.5 M TCEP-HCI (Tris(2-Carboxyethyl)Phosphine Hydrochloride) in high purity dH2O.

pH (25°C): 6.6 ± 0.1Concentration (TCEP): 0.5 - 0.55 M

77531 5 ml 50 ml

TE Buffer, 1X SolutionULTRAPURE

MB GradeForm: 1X TE solution consisting of 10 mM Tris, pH

7.5, and 1 mM EDTA. Solution is 0.2 µm filtered and dispensed into vials and/or durable, square Nalgene bottles.

pH (25°C): 7.5 ± 0.1Contaminant Testing:


Application: � Use directly to resuspend and/or dilute purified

DNA or RNA.

75893 10 x 1 ml 100 ml 500 ml

TE Buffer, 1X Solution, pH 7.0

Form: 1X TE solution consisting of 10 mM Tris Ultrapure, pH 7.0, and 1 mM EDTA. Solution is 0.2 μm filtered and dispensed into vials and/or durable, square Nalgene bottles.

pH (25°C): 7.0 ± 0.1 Contaminant Testing:


Application:� Use directly to re-suspend and/or dilute purified

DNA or RNA.

75790 100 ml 500 ml

TE Buffer, 1X Solution, pH 7.0, Low EDTA

Form: 1X TE solution consisting of 10 mM Tris Ultrapure, pH 7.0, and 0.1 mM EDTA. Solution is 0.2 μm filtered and dispensed into vials and/or durable, square Nalgene bottles.




DNA or RNA.

75791 100 ml 500 ml

TE Buffer, 1X Solution, pH 8.0

MB GradeForm: 1X TE solution consisting of 10 mM Tris

Ultrapure, pH 8.0, and 1 mM EDTA. Solution is 0.2 μm filtered and dispensed into vials and/or durable, square Nalgene bottles.




DNA or RNA.

75792 100 ml 500 ml

TE Buffer, 1X Solution, pH 8.0, Low EDTA

MB GradeForm: 1X TE solution consisting of 10 mM Tris

Ultrapure, pH 8.0, and 0.1 mM EDTA. Solution is 0.2 μm filtered and dispensed into vials and/or durable, square Nalgene bottles.




DNA or RNA.

75793 100 ml 500 ml

TE Buffer, 50X SolutionULTRAPURE

Form: 50X TE solution consisting of 500 mM Tris, 50 mM EDTA in high purity dH2O, adjusted to pH 7.5. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. Dilute to 1X with high purity dH2O for a solution consisting of 10 mM Tris, pH 7.5, and 1 mM EDTA.

pH (25°C, 50X): 7.5 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes test

75834 100 ml 500 ml

Biochemicals


TEMED

See Tetramethylethylenediamine, page 245

Terrific Broth, Ready-Made Powder

Form: Off-white free flowing powder.pH (5.08%, H2O): 7.2 ± 0.2Contents: Casein Peptone: 12 gm/L Yeast Extract: 24 gm/L Potassium Phosphate Dibasic: 12.5 gm/L Potassium Phosphate Monobasic: 2.3 gm/LApplication: � Provides greater yields of plasmid DNA and

recombinant proteins than LB media by allowing for greater cell density.

To use: Add 50.8 gm powder, 4 ml glycerol, and 996 ml water to make 1 liter medium. Dissolve and autoclave.

75856 250 gm 1 kg

TES(N-Tris[Hydroxymethyl]methyl-2-Aminoethanesulfonic Acid)

ULTRAPURE

C6H15NO6SFormula Wt.: 229.25Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity A: KBr pellet matches referenceIdentity B: Passes testpKa (20°C): 7.50 ± 0.15A (250 nm, 1 M, 1 cm): ≤ 0.2Loss on Drying: ≤ 1.0%Iron (Fe): ≤ 30 ppmLead (Pb): ≤ 10 ppmCAS #7365-44-8

22086 100 gm 500 gm 1 kg

Tetracycline, HydrochlorideULTRAPURE


•HClFormula Weight: 480.90Product specifications:Form: Yellow crystalline powderActivity: ≥ 900 µg of C22H24N2O8

•HCI/mg (as is)Identity (A, B, C, D, E, F): Passes testsCrystallinity: Passes testpH (10 mg/ml): 1.8 - 2.8[α]25

D : -240° to -255° (c=0.5, 0.1N HCI, dry basis)Loss on Drying (60°C, 3 hours, vacuum): ≤ 2.0%Limit of 4-Epianhydrotetracycline: ≤ 2.0%Heavy Metals (Pb): ≤ 0.005%Storage: Ambient in tightly closed, light-resistant

containers.CAS #64-75-5

22105 25 gm 100 gm 1 kg

Tetramethylammonium Chloride(TMAC)

ULTRAPURE

C4H12ClNFormula Weight: 109.60Product specifications:Form: White crystalline powderAssay: ≥ 98% (dry basis)Identity: By IRpH (10%, H2O): ≥ 4.5Loss on Drying: ≤ 1.5%Solubility (10%, H2O): Clear and colorlessApplications:� Used in hybridization solutions of mixed oligo-

nucleotides. In solutions with TMAC, the Tm (melt-ing temperature) of a hybrid is dependent on its length, not its base composition.

� Used in probing PCR products for the analysis of polymorphisms with allele-specific oligonucle-otides.

Storage: Store at room temperature. This material is extremely hygroscopic.

CAS #75-57-0

22124 1 kg

3,3',5,5'-Tetramethylbenzidine(TMB)

C16H20N2

Formula Weight: 240.35Product specifications:Form: Off-white powderIdentity: By IRMelting Point: 168.3 - 169.9°CCAS #54827-17-7

22126 1 gm 5 gm

N,N,N',N'-Tetramethylethylene diamine(TEMED)

ULTRAPURE

MB Grade(CH3)2NCH2CH2N(CH3)2

Formula Weight: 116.20Product specifications:Form: Colorless to pale yellow liquidAssay: ≥ 98%A (290 nm, 1%, 1 cm): ≤ 0.1Density (20°C): 0.77 ± 0.01Refractive Index (20°C): 1.419 ± 0.001Testing: Meets or exceeds functional test criteria for

high quality gel electrophoresis.Application: � Catalyzes the formation of free radicals by ammo-

nium persulfate and accelerates the polymeriza-tion of acrylamide and bisacrylamide.

CAS #110-18-9.

76320 100 ml 100 gm 500 gm

Tetrazolium Red

See 2,3,5-Triphenyl Tetrazolium Chloride, page 246

Thiamine, Hydrochloride(Vitamin B1 ; Aneurine)

USBioAnalyzedC12H17CIN4OS•HClFormula Weight: 337.27Product specifications:Form: White crystalline powderAssay: 98.0 - 102.0% (dry basis)Identity (A, B): Passes testsChromatographic Purity: Passes testpH (1%, H2O): 2.7 - 3.4Spectral Ratios (400 nm, 10%, 1 cm): ≤ 0.025Moisture (KF): ≤ 5.0%Residue on Ignition: ≤ 0.2%Nitrate: Passes testResidual Solvents: Meets the requirementsCAS #67-03-8

22170 100 gm 500 gm 1 kg

Thiamine Pyrophosphate Chloride

See Cocarboxylase, page 207

Thiazolyl Blue

See MTT, page 228

Biochemicals


Thimerosal

USBioAnalyzedC9H9HgNaO2SFormula Weight: 404.81Product specifications:Form: White powderAssay: 97.0 - 101.0% (dry basis)Identity (A, B): Passes testsLoss on Drying: ≤ 0.5%Ether-Soluble Substances: Passes testMercury Ions: ≤ 0.70%Readily Carbonizable Substances: Passes testStorage: Store in light-resistant containers.CAS #54-64-8

22215 25 gm 100 gm

Thioredoxin, Human, Recombinant(r-hTRX)

Formula Weight: 11,734 determined by amino acid sequence

Product specifications:Form: Lyophilized powder from phosphate bufferSpecific Activity: ≥ 3 units/mg proteinUnit Definition: TRX activity is assayed by measuring

the change in absorbance at 650 nm at 25°C using 0.13 µM bovine insulin containing 0.33 mM DTT (pH 6.5).

Storage: -20°C, protect from moisture.

26004 100 µg

L-Threonine

CH3CH(OH)CH(NH2)COOHFormula Weight: 119.12Product specifications:Form: White crystalline powderAssay: ≥ 98.5%Identity: By IRpH (5%, H2O): 5.0 - 6.5[α]25

D : -26.7° to -29.1° (c=6 in H2O)Loss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.4%Heavy Metals (Pb): ≤ 15 ppmChloride (Cl): ≤ 0.05%Iron (Fe): ≤ 30 ppmSulfate (SO4): ≤ 0.03%Storage: Store in light-resistant containers.CAS #72-19-5

22290 100 gm 1 kg

Thrombin Human

Source: Human1,000 NIH units/vialProduct specifications:Form: Lyophilized powderActivity: ≥ 1,000 NIH units/vialSpecific Activity: ≥ 2,000 NIH units/mg proteinReconstitution: Reconstitute with 1.0 ml ddH2OBuffer: After reconstitution the solution contains: 5 mM Sodium Citrate 0.01% PEG 0.02 M NaCl pH 6.5All source material tested and found non-reactive

to HTLV-III and HBsAg using FDA approved assay methods.

Storage: -20°C

22292 1,000 units

Thrombin

(E.C.3.4.21.5)Source: Bovine BloodProduct specifications:Form: Lyophilized powderActivity: ≥ 40 units/mg powderMoisture: ≤ 5.0%Note: The activity is determined by comparison

to the activity of Standard Thrombin from the Bureau of Biological Standards.

Storage: 4°C, in tightly closed containers.CAS #9002-04-4

22293 25,000 units 100,000 units

TMAC

See Tetramethylammonium Chloride, page 245

TMB

See 3,3',5,5'-Tetramethylbenzidine, page 245

Toxillic Acid

See Maleic Acid, page 226

α,α-Trehalose, Dihydrate(α-D-Glucopyranosyl-α-D-Glucopyranoside)

C12H22O11•2H2O

Formula Weight: 378.33Product specifications:Form: White crystalline powderMoisture (KF): 9.3 - 9.7%[α]20

D : +179.0° ± 1.5° (c=7, H2O)Arsenic (As): ≤ 0.5 ppmIron (Fe): ≤ 5 ppmHeavy Metals (Cu): ≤ 10 ppmCAS #6138-23-4

22515 100 gm

Tricine[N-Tris (Hydroxymethyl)methylglycine]

ULTRAPURE

(CH2OH)3CNHCH2COOHFormula Weight: 179.17Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (20°C): 8.15 ± 0.15A (250 nm, 5%, 1 cm): ≤ 0.1Loss on Drying: ≤ 0.5%Iron (Fe): ≤ 30 ppmLead (Pb): ≤ 10 ppmCAS #5704-04-1

22561 100 gm 500 gm 1 kg

Triethanolamine, Hydrochloride

N(CH2CH2OH)3•HCl

Formula Weight: 185.65Product specifications:Form: White/clear crystalline powderAssay: ≥ 98.5% (dry basis)Identity: By IRpKa (20°C): 7.85 ± 0.15Melting Point: 178 - 182°CLoss on Drying: ≤ 1%Iron (Fe): ≤ 30 ppmHeavy Metals (Pb): ≤ 10 ppmCAS #637-39-8

22565 250 gm 1 kg

1,3,7-Trimethylxanthine

See Caffeine, page 204

2,6,8-Trioxypurine

See Uric Acid, page 249

2,3,5-Triphenyl Tetrazolium Chloride(TTC; Tetrazolium Red)

C19H15ClN4

Formula Weight: 334.80Product specifications:Form: Cream to beige colored powderIdentity: By IRSolubility (1%, H2O): Clear solutionMoisture: ≤ 5% (By KF)Sensitivity for Reducing Sugars: Passes testCAS #298-96-4

22631 10 gm 25 gm 50 gm 100 gm 500 gm 1 kg

Biochemicals


Triphosphopyridine Nucleotide Dinucleotide

See Nicotinamide Adenine Dinucleotide Reduced, page 230

2,4,6-Tripyridyl-s-Triazine (TPTZ)

C18H12N6

Formula Weight: 312.33CAS #3682-35-7

22660 5 gm 25 gm 100 gm

Tris(Tris[hydroxymethyl]aminomethane)

ULTRAPURE

MB GradeNH2C(CH2OH)3

Formula Weight: 121.14Product specifications:Form: White crystalline powderAssay: 99.8 - 100.1% (dry basis)Identity: By IRpKa (20°C): 8.30 ± 0.15A (250 nm, 1 M, 1 cm): ≤ 0.1A (290 nm, 40%, 1 cm): ≤ 0.2Solubility (1 M, 500 ml): Clear and colorlesspH (5%, H2O): 10.0 - 11.5Melting Point: 168 - 172°CLoss on Drying: ≤ 1%Moisture (KF): ≤ 1%Residue on Ignition: ≤ 0.1%Insoluble Matter: ≤ 0.005%Arsenic (As): ≤ 1 ppmBarium (Ba): ≤ 1 ppmCadmium (Cd): ≤ 1 ppmCalcium (Ca): ≤ 1 ppmCopper (Cu): ≤ 1 ppmIron (Fe): ≤ 1 ppmLead (Pb): ≤ 1 ppmMagnesium (Mg): ≤ 1 ppmManganese (Mn): ≤ 1 ppmZinc (Zn): ≤ 1 ppmDNase (endo): Passes testDNase (exo): Passes testRNase: Passes testProtease: Passes testCAS #77-86-1

75825 500 gm 1 kg 5 kg 10 kg

Tris[Tris(hydroxymethyl)aminomethane]

High Purity GradeNH2C(CH2OH)3

Formula Weight: 121.14Product specifications:Form: White/clear crystalline powderAssay: 99.0 - 101.0% (dry basis)Identity: By IRpKa (20°C): 8.30 ± 0.15A (250 nm, 1 M, 1 cm): ≤ 0.1A (290 nm, 40%, 1 cm): ≤ 0.2pH (5%, H2O): 10.0 - 11.5Melting Point: 168 - 178°CLoss on Drying: ≤ 1%Residue on Ignition: ≤ 0.1%Insoluble Matter: ≤ 0.005%Iron (Fe): ≤ 5 ppmHeavy Metals (Pb): ≤ 5 ppmCAS #77-86-1

22674 500 gm 1 kg 5 kg

Tris[Tris(hydroxymethyl)aminomethane]

Reagent GradeNH2C(CH2OH)3

Formula Weight: 121.14Product specifications:Form: White to pale yellow crystalline powderAssay: ≥ 98.0% (dry basis)Identity: By IRpH (5%, H2O): 10.0 - 11.5Melting Point: ≥ 160°CLoss on Drying: ≤ 1.5%CAS #77-86-1

22675 500 gm 1 kg

Tris-Glycine (TG) Buffer, 10X SolutionULTRAPURE

MB GradeForm: 10X Tris-Glycine (TG) Buffer consisting of

0.25 M Ultrapure Tris, pH 8.3, and 1.92 M gly-cine. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout. Dilute to 1X with high purity water and methanol for a solution consisting of 25 mM Tris, pH 8.3, 192 mM glycine, 20% methanol.

Contaminant Testing: Protease: Passes test.

Application: � For Western blotting and gel electrophoresis.

75894 1 L 5 L

Tris-Glycine-SDS (TGS) Buffer, 10X SolutionULTRAPURE

MB GradeForm: 10X Tris-Glycine-SDS (TGS) Buffer, consisting

of 0.25 M Ultrapure Tris, pH 8.6, 1.92 M glycine, and 1.0% SDS. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout. Dilute to 1X with high purity dH2O for a solution consisting of 25 mM Tris, pH 8.6, 192 mM glycine and 0.1% SDS.

Contaminant Testing: Protease: Passes testApplication: � TGS is the most commonly used buffer for SDS-

PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) of proteins.

75895 1 L 5 L

Tris, Hydrochloride(Tris[hydroxymethyl]aminomethane Hydrochloride)

ULTRAPURE

MB GradeC4H11NO3 HClFormula Weight: 157.60Product specifications:Form: White crystalline powderAssay: 99.0 - 101.0% (as is)Identity: By IRpKa (20°C): 8.30 ± 0.15A (250 nm, 1 M, 1 cm): ≤ 0.20A (290 nm, 40%, 1 cm): ≤ 0.20pH (5%, H2O): 4.7 ± 0.5Melting Point: 148 - 157°CLoss on Drying: ≤ 1.0%Residue on Ignition: ≤ 0.1%Insoluble Matter: ≤ 0.005%Iron (Fe): ≤ 5 ppmHeavy Metals (Pb): ≤ 5 ppmDNase (endo): Passes testDNase (exo): Passes testRNase: Passes testProtease: Passes testStorage: Store in tightly closed containers.CAS #1185-53-1

22676 500 gm 1 kg 5 kg

Biochemicals


Tris/HCl, 1 M Solution, pH 7.0ULTRAPURE

MB GradeForm: 1 M solution of Tris in high purity dH2O,

adjusted to pH 7.0. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles.

pH (25°C): Passes testContaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testCAS #1185-53-1

22637 100 ml 500 ml 1 L


MB GradeForm: 1 M solution of Tris in high purity dH2O,

adjusted to pH 7.5. Solution is 0.2 µm filtered and dispensed into durable, square Nalgene bottles.

pH (25°C): 7.5 ± 0.1Contaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes testCAS #1185-53-1

22639 100 ml 500 ml 1 L


MB GradeForm: 1 M solution of Tris, Ultrapure in high purity

dH2O, adjusted to pH 8.0. Solution is 0.2 µm fil-tered and dispensed into durable, square Nalgene bottles.

pH (25°C): Passes testContaminant Testing: DNase (endo): Passes test DNase (exo): Passes test RNase: Passes test Protease: Passes test

22638 100 ml 500 ml 1 L

r-hTRX

See Thioredoxin, Human, Recombinant, page 246

TrypsinULTRAPURE

(E.C.3.4.21.4)USBioAnalyzedSource: Bovine PancreasProduct specifications:Form: White to pale yellow powderActivity: ≥ 2,500 USP units/mg powderContaminating Activity:

Chymotrypsin: ≤ 50 USP units/mg powderLoss on Drying: ≤ 5.0%Solubility: Passes testResidue on Ignition: ≤ 2.5%Microbial Limits Test per USP: Passes testStorage: -20°C, protect from moisture.CAS #9002-07-7

22720 1 gm 5 gm 10 gm

Trypsin (TPCK Treated)(E.C.3.4.21.4)Source: Bovine PancreasProduct specifications:Form: Lyophilized powder, treated with TPCK to

inhibit chymotryptic activityActivity: ≥ 180 units/mg proteinUnit Definition: One unit hydrolyzes 1 µmol of

TAME/minute at 25°C, pH 8.2, in the presence of 0.01 M Ca2+.


22725 100 mg 250 mg

Trypsin 1:250

(E.C.3.4.21.4)Source: Porcine PancreasProduct specifications:Form: Powder stabilized with lactose.Activity: ≥ 250,000 USP units/gmChymotrypsin: ≥ 75,000 USP units/gmUnit Definition: One unit causes a change in absor-

bance of 0.003/minute with BAEE as a substrate at 25°C, pH 7.6.


22710 100 gm 1 kg

L-Tryptophan(L-2-Amino-3-Indole Propionic Acid)


Formula Weight: 204.23Product specifications:Form: White to off-white crystalline powderAssay: 98.5 - 101.5% C11H12N2O2 (dry basis)Identity: By IRpH (1%, H2O): 5.5 - 7.0Chromatography (TLC): Homogeneous[α]25

D : -29.4° to -32.8° (c=1 in H2O)Loss on Drying: ≤ 0.3%Residue on Ignition: ≤ 0.1%Heavy Metals (Pb): ≤ 0.0015%Chloride (Cl): ≤ 0.05%Iron (Fe): ≤ 0.003%Sulfate (SO4): ≤ 0.03%Residual Solvents: Meets the requirementsCAS #73-22-3

22765 100 gm 1 kg

Tyrosinase(Polyphenol Oxidase)

(E.C.1.14.18.1)Source: MushroomProduct specifications:Form: Lyophilized powderActivity: ≥ 500 units/mg (dry basis)Storage: -20°C

22885 100,000 units 500,000 units

L-Tyrosine

USBioAnalyzedHO(C6H4)CH2CH(NH2)COOHFormula Weight: 181.19Product specifications:Form: White to off-white crystalline powderAssay: 98.5 - 101.5% (dry basis)Identity: By IR[α]25

D : -9.8° to -11.2° (dry basis, c=5 in 1N HCl)Loss on Drying (105°C, 3 hours): ≤ 0.3%Residue on Ignition: ≤ 0.4%Chloride (Cl): ≤ 0.04%Iron (Fe): ≤ 30 ppmSulfate (SO4): ≤ 0.04%Heavy Metals (Pb): ≤ 15 ppmChromotographic Purity: Passes testResidual Solvents: Meets the requirementsCAS #60-18-4

22910 100 gm 1 kg 5 kg

Biochemicals


Uracil

C4H4N2O2

Formula Weight: 112.09Product specifications:Form: White to off-white crystalline powderAssay (Total UV, pH 7.0): ≥ 98.0% (dry basis)Identity: By IRSpectral Ratios (pH 7.0):

A280/A260: 0.18 ± 0.02 A250/A260: 0.84 ± 0.03Loss on Drying (105°C, 2 hours): ≤ 5.0%Residue on Ignition: ≤ 0.1%Reference constants:λmax (pH 7, NRC): 260 nmεmax (pH 7, NRC): 8.2 x 103

ε260 (pH 7, DBR): 8.2 x 103

CAS #66-22-8

23020 100 gm 1 kg

UreaULTRAPURE

MB GradeNH2CONH2

Formula Weight: 60.06Product specifications:Form: White crystalline powderAssay: ≥ 99.5%Identity: By IRSpectral Ratios (260 nm, 6 M, 1 cm): ≤ 0.055Spectral Ratios (280 nm, 6 M, 1 cm): ≤ 0.044Melting Point: 132 - 135°CInsoluble Matter ≤ 0.01%Residue on Ignition: ≤ 0.01%Conductivity: ≤ 15 µmhos/cmCyanate: None detectedChloride (Cl): ≤ 0.0005%Sulfate (SO4): ≤ 0.001%Copper (Cu): ≤ 0.5 ppmIron (Fe): ≤ 0.5 ppmLead (Pb): ≤ 0.5 ppmDNase (endo): Passes testRNase: Passes testProtease: Passes testApplications:� Chaotropic agent used in denaturing polyacryla-

mide gel electrophoresis.� When 8 M urea is not sufficiently denaturing,

40% formamide may be included.CAS #57-13-6

75826 1 kg 5 kg

Urea

High Purity GradeNH2CONH2

Formula Weight: 60.06Product specifications:Form: White crystalline powderAssay: 99.0 - 100.5%Identity: By IRLoss on Drying: ≤ 1.0%Melting Point: 132 - 135°CInsoluble Matter: ≤ 0.01%Residue on Ignition: ≤ 0.01%Chloride (Cl): ≤ 5 ppmSulfate (SO4): ≤ 0.001%Heavy Metals (Pb): ≤ 0.001%Iron (Fe): ≤ 0.001%CAS #57-13-6

23036 1 kg 5 kg

Uric Acid(2,6,8-Trioxypurine)

C5H4N4O3

Formula Weight: 168.11Product specifications:Form: White to off-white powderAssay: ≥ 98.5%Identity: By IRSolubility: Clear solutionLoss on Drying: ≤ 0.2%Residue on Ignition: ≤ 0.2%Heavy Metals: ≤ 20 ppmCAS #69-93-2

23075 100 gm 500 gm 1 kg

Uridine

C9H12N2O6

Formula Weight: 244.20Product specifications:Form: White to slightly off-white powderAssay (UV): 100% ± 2%Identity: By IRλmax (pH 7.0): 262 nm ± 1 nmεmax (pH 7.0): [10.1 ± 2%] x 103


A280/A260: 0.36 ± 0.02Loss on Drying: ≤ 1.0%Heavy Metals (Pb): ≤ 10 ppmReference constants:λmax (pH 7, NRC): 262 nmεmax (pH 7, NRC): 10.1 x 103

ε260 (pH 7, DBR): 10.0 x 103

CAS #58-96-8

23130 1 gm 10 gm 100 gm

Uridine-5'-Triphosphate, Trisodium Salt(UTP)


Formula Weight: 586.12Product specifications:Form: White crystalline powderAssay (Total UV): 100% ± 3% (dry basis)Assay (HPLC): ≥ 90%λmax (pH 7.0): 262 nm ± 1 nmεmax (pH 7.0): [10.0 ± 0.3] x 103

Spectral Ratios (pH 7.0):A250/A260: 0.60 - 0.90

A280/A260: 0.30 - 0.45Moisture: ≤ 15.0%Reference constants:λmax (pH 7, NRC): 262 nmεmax (pH 7, NRC): 10.0 x 103

ε260 (pH 7, DBR): 9.9 x 103

Storage: -20°CCAS #19817-92-6

23160 100 mg 1 gm

UTP

See Uridine-5'-Triphosphate, page 249

Biochemicals


Vancomycin, Hydrochloride

USBioAnalyzedC66H75Cl2N9O24

•HClFormula Weight: 1485.71Product specifications:Form: Tan to brown powderActivity: ≥ 900 µg of vancomycin/mg (dry basis) ≥ 855 µg of vancomycin/mg (as is)Identity: By IRChromatographic Purity: Vancomycin B: ≥ 80.0% Other Peaks: ≤ 9%pH (50 mg/ml, H2O): 2.5 - 4.5Moisture (KF): ≤ 5.0%Storage: 4°C, in tightly closed containers.CAS #1404-93-9

23261 100 mg 250 mg

Vitafree Casein

See Casein Vitafree, page 205

Vitamin B1

See Thiamine HCl, page 245

Vitamin B2

See Riboflavin, page 238

Vitamin B5

See D-Calcium Pantothenate, page 204

Vitamin B12

(Cyanocobalamin)

USBioAnalyzedC63H88CoN14O14PFormula Weight: 1355.37Product specifications:Form: Dark red crystalline powderAssay: 96.0 - 100.5% (dry basis)Identification A: Passes testIdentification B: Passes testIdentification C: Passes testLoss on Drying: ≤ 12.0%Psuedocyanocobalamin: Passes testStorage: In tightly closed, light-resistant containers.CAS #68-19-9

23360 1 gm

Vitamin C

See L-Ascorbic Acid, page 201

Vitamin Free Casein

See Casein-Vitafree, page 205

Vitamin G

See Riboflavin, page 238

Vitamin K1

(Phytonadione; 2-methyl-3-Phytyl-3-1; 4-Napthoquinone)


Formula Weight: 450.70Product specifications:Form: Clear yellow to amber viscous liquidAssay: 97.0 - 103.0%Identity (A, B): Passes testsRefractive Index (25°C): 1.523 to 1.526Reaction: Neutral to litmusLimit of Menadione: Passes testZ-isomer content: ≤ 21.0%CAS #84-80-0

23445 250 mg 1 gm 5 gm 10 gm

Water, Nuclease-FreeULTRAPURE

MB GradeForm: High purity dH2O tested for contaminating

nuclease activity. Product is 0.2 µm filtered and dispensed into vials and/or durable, square Nalgene bottles. The 5 L size is supplied in an upright carton with a convenient dispensing spout.

Contaminant Testing: RNase: Passes test DNase (endo): Passes test DNase (exo): Passes test Protease: Passes testAssays: Functional Assay: Functionally tested for PCR

according to the standard Affymetrix PCR protocol.

Enzyme Inhibition Assay: No inhibition by nuclease-free H2O on digestion of lambda DNA by Hind III.

71786 10 x 1 ml 100 ml 500 ml 1 L 5 L

Water, RNase-Free, DEPC TreatedULTRAPURE

MB GradeForm: High purity dH2O treated overnight with 0.1%

Diethylpyrocarbonate. Product is 0.2 µm filtered and dispensed into vials and/or durable, square Nalgene bottles, then autoclaved.

Contaminant Testing: RNase: Passes test DNase (endo): Passes test DNase (exo): Passes test Protease: Passes testAssays: Functional Assay: Functionally tested for PCR

according to the standard Affymetrix PCR protocol.

Enzyme Inhibition Assay: No inhibition by RNase-free H2O on digestion of lambda DNA by Hind III.

70783 25 ml 10 x 1 ml 100 ml 500 ml 1 L

Wright’s Stain

Prepared for use in blood stain work.CAS #68988-92-1

23457 5 gm 10 gm 25 gm 100 gm

Xanthine Monosodium Salt, Monohydrate

C5H3N4O2Na•H2OFormula Weight: 192.11Product specifications:Form: White to off-white crystalline powderAssay (Total UV): ≥ 96.0% (dry basis)Identity: By IRMoisture (KF): ≤ 12.0%Spectral Ratios (pH 10.0): A250/A260: 1.29 ± 0.06 A280/A260: 1.71 ± 0.05Reference constants:λmax (pH 10, NRC): 277 nmεmax (pH 10, NRC): 9.3 x 103

ε260 (pH 10, DBR): 5.2 x 103

CAS #1196-43-6

23465 5 gm 10 gm 25 gm 100 gm

Biochemicals


X-α-GAL

See 5-Bromo-4-Chloro-3-Indolyl-α-D-Galactopyranoside, page 203

X-Gal

See 5-Bromo-4-Chloro-3-Indolyl-β-D-Galactopyranoside, page 203

Xylene Cyanol FFULTRAPURE

(C.I.43535)C25H27N2NaO7S2

Formula Weight: 554.61Product specifications:Form: Dark burgundy metallic powderIdentity: By IRεmax (615 nm, H2O, 1 cm): ≥ 60 x 103

Loss on Drying: ≤ 10.0%λmax (H2O solution): 615 nm ± 2 nmApplication:� Useful as a tracking dye during the separation of

nucleic acids (in particular, in DNA sequencing electrophoresis).

Sequencing Stop Solution: 95% Formamide 20 mM EDTA 0.05% Bromophenol Blue 0.05% Xylene Cyanol FF Store solution at -20°C.10X Agarose Gel Loading Solution: 50% Glycerol 60 mM EDTA 0.05% Bromophenol Blue 0.05% Xylene Cyanol 1.0% SDSStorage: Store solution at room temperature.CAS #4463-44-9

23513 25 gm

D-(+)-Xylose

C5H10O5

Formula Weight: 150.13Product specifications:Form: White crystalline powderAssay: ≥ 98.5%[α]20

D : +18.9° to +19.7° (c=4 in H2O)Loss on Drying: ≤ 1.0%Residue on Ignition: ≤ 0.05%Heavy Metals (Pb): ≤ 5 ppmCAS #58-86-6

23520 500 gm 1 kg

Yeast, Bakers

Source: Baker’s YeastStorage: 4°C

23540 1 lb 5 lb 25 lb

Yeast, Brewers

CAS #68876-77-7

23546 5 lb 25 lb 100 lb

Yeast Extract Powder(Peptones)

ULTRAPURE

Source: Baker’s YeastProduct specifications:Form: Off-white powderProtein (TN x 6.25): 57-72%Moisture: ≤ 6% at time of packagingSalmonella: NegativeStandard Plate Count: ≤ 15,000/gmThis product is hygroscopic.Storage: Ambient, in tightly closed containers.CAS #8013-01-2

23547 1 kg

Yeast Extract Powder, Low Dust (Peptones)

Source: Baker’s YeastProduct specifications:Form: Tan powder, low dustProtein (TN x 6.25): 57 - 72%Moisture: ≤ 6% at time of packagingSalmonella: NegativeStandard Plate Count: ≤ 15,000/gmThis product is hygroscopic.Storage: Ambient, in tightly closed container.CAS #8013-01-2

23545 1 kg 5 kg

Yeast Hydrolysate, Enzymatic(Peptones)

ULTRAPURE

Product specifications:Form: Light tan powderIdentity: By IRSolubility (5%, H2O): Passes testpH (5% H2O): 5.4 - 6.2CAS #100684-36-4

23550 5 lb 25 lb 50 lb 100 lb

Yeast-Torula

23551 5 lb 25 lb 100 lb

2X YT Broth

Form: Off-white, free-flowing powderpH (3.1%, H2O): 7.0 ± 0.2Contents: Casein Peptone: 16 gm/L Yeast Extract: 10 gm/L Sodium Chloride: 5 gm/LTo Use: Dissolve 31 gm powder per liter of purified

water.

75864 1 kg

Catalog IndexProduct Code Index

Product Name Index

Code Product Size Page Code Product Size Page

Product Code Index | Numerical


10062 S-AcetylCoenzymeA,LithiumSalt,Ultrapure 10mg 188 25mg 100mg 500mg

10077 5-Bromo-4-Chloro-3-Indolyl-b-D-Galactopyranoside, 100mg 203 Ultrapure 250mg 1gm 2gm 5gm

10120 N-Acetyl-L-Cysteine 100gm 188 500gm 1kg

10132 Agarose-Hi-Res,Separation≤1000bp,Ultrapure 25gm 195 100gm 500gm

10430 Adenine 100gm 188 1kg

10450 AdenineSulfate,Dihydrate 100gm 188 1kg

10460 Adenosine 100gm 189

10490 Adenosine-5’-Diphosphate,DisodiumSalt,Dihydrate 5gm 189 10gm 25gm

10520 Adenosine-5’-Monophosphate,FreeAcid 10gm 189 25gm 100gm

10521 Adenosine-5’-Monophosphate,FreeAcid,Monohydrate 25gm 189 100gm 1kg

10585 Adenosine-5’-Triphosphate,DisodiumSalt 5gm 189 10gm 25gm 100gm 500gm

10601 S-Adenosyl-L-MethionineSulfatep-Toluenesulfonate 25mg 190 100mg

10620 Adenosine-5’-Monophosphate,DisodiumSalt 25gm 189 100gm

10635 Adonitol 25gm 190 100gm

10654 Agar 1lb 190 5lb 25lb

10665 b-Alanine 200gm 198 1kg 5kg

10848 Albumin,Bovine,Acetylated,Ultrapure 25mg 198 125mg

10852 Albumin,Bovine,CohnFractionV,pH5.2 50gm 198 100gm 500gm

10856 Albumin,Bovine,Crystallized,pH5.2 5gm 198 10gm 100gm

10857 Albumin,Bovine,pH7 50gm 198 100gm 500gm 1kg

10865 Albumin,Egg 5gm 199

10867 Albumin,Bovine,Protease-Free 10gm 198 50gm 100gm

10868 Albumin,Bovine,RIAGrade,pH7 10gm 198 50gm 100gm

10895 AlcoholDehydrogenase 75,000units 199

10906 Agar,Bacteriological,Ultrapure 1lb 190 5lb 25lb

10907 Agar,NobleAgar,Ultrapure 100gm 190 500gm 1kg

10925 ChickenIntestineAlkalinePhosphastase 1gm 206 5gm

11055 p-AminoBenzoicAcid(PABA) 200gm 199 500gm 1kg 5kg

11056 4-AminoAntipyrene 50gm 199 100gm

11079 L-a-Amino-N-ButyricAcid 1gm 199 5gm

11118 AEBSFHydrochloride 200mg 190 1gm

11251 AmmoniumAcetate,Ultrapure 500gm 199 1kg

11254 AmmoniumSulfate,Ultrapure 1kg 200 5kg

11259 Ampicillin,SodiumSalt,Ultrapure 5gm 200 25gm 100gm

11379 G-418Sulfate,Ultrapure 500mg 217 1gm 5gm

11388 Aprotinin,Ultrapure 10mg 200 25mg 100mg

11405 L-(+)-Arabinose 25gm 200 100gm 1kg

11406 L-Arabinose 25gm 200 100gm 1kg

11490 L-Arginine 100gm 200 500gm 1kg

11500 L-Arginine,Hydrochloride 100gm 201 500gm 1kg

11545 L-AscorbicAcid 1kg 201 5kg

11590 L-Asparagine,Anhydrous 100gm 201 500gm 1kg




11595 L-Asparagine,Monohydrate 100gm 201 1kg 5kg

11620 L-AsparticAcid 100gm 201 1kg 5kg

11631 L-AsparticAcid,MagnesiumSalt,Dihydrate 100gm 201 1kg

11635 L-AsparticAcid,SodiumSalt,Monohydrate 1kg 201

11681 Streptavidin,Ultrapure 5mg 242 10mg

11687 Streptavidin-AlkalinePhosphataseConjugate 250µl 242 1,500µl

11805 Bacitracin 5gm 202 25gm

12091 Bicine 100gm 202 1kg

12110 Bilirubin 1gm 202 5gm

12112 Bis-Tris,Ultrapure 100gm 202 500gm 1kg

12355 5-Bromo-4-Chloro-3-IndolylBromocresolGreen, 5gm 203 SodiumSalt,Ultrapure 25gm

12370 BromophenolBlue,SodiumSalt,Ultrapure 5gm 204 10gm

12387 5-Bromo-4-Chloro-3-5-Bromo-4-Chloro-3-Indolyl 100mg 203 Phosphate,DisodiumSalt,Ultrapure 500mg 1gm

12488 CacodylicAcid,SodiumSaltTrihydrate 100gm 204 500gm

12520 Caffeine,Anhydrous,Ultrapure 100gm 204 1kg

12531 CalciumChloride,Dihydrate 500gm 204 2.5kg

12550 D-CalciumPantothenate 100gm 204 500gm 1kg

12678 Carbenicillin,DisodiumSalt 1gm 205 5gm

12758 CarboxypeptidaseY 1mg 205 10mg

12840 Casein,Hammarsten,Ultrapure 100gm 205 1lb 5lb 25lb

12845 Casein(HighNitrogen) 5lb 205 25lb 100lb

12855 CaseinHydrolysate,EnzymaticDigest,Ultrapure 1lb 205 5lb 25lb

12865 Casein,Sodium 1lb 205 5lb

12866 Casein-Vitafree,Ultrapure 5lb 205 25lb

12885 Catalase 500mg 205 1gm 5gm 10gm

13285 Cellulase 100gm 205

13353 CetylTrimethylAmmoniumBromide 100gm 206 500gm 1kg

13410 CholineChloride 200gm 207 1kg 5lb 25lb

13580 Cholesterol,ReagentGrade 5gm 206 100gm

13610 Cholesterol 100gm 206 1kg

13729 CitricAcid,Anhydrous 500gm 207 1kg

13735 CitricAcid,TrisodiumSalt,Dihydrate,Ultrapure 1kg 207 5kg

13765 Cocarboxylase 25gm 207 100gm

13785 CoenzymeA,TrilithiumSalt,Dihydrate 100mg 207 250mg 500mg 1gm

13787 CoenzymeA,FreeAcid,Trihydrate 50mg 207 100mg 250mg 500mg 1gm

13813 Collagen 10ml 207 25ml 100ml

13820 Collagenase,TypeI 100mg 207 1gm 5gm

13911 CRCLowerFilters,10µmporesize 1pk 208



13914 CRCUpperFilters,10µmporesize 1pk 208

13915 CreatineKinase 100mg 208 1gm



13920 CreatinePhosphate,DisodiumSalt,Tetrahydrate 5gm 208 25gm

13928 CompactReactionColumns 1pk 207

13929 CompactReactionColumns,withoutscrewonlids 1pk 208

13979 a-Cyclodextrin 25gm 208




14035 L-Cysteine,Hydrochloride,Monohydrate 25gm 208 100gm 1kg 5kg

14050 L-Cystine 50gm 208 100gm 1kg 5kg

14060 Cytidine,FreeBase 25gm 208 100gm

14102 CytochromeC 100mg 209 500mg 1gm

14121 Cytidine-5’-Triphosphate,DisodiumSalt,Dihydrate 100mg 209 500mg 1gm

14235 2’-Deoxyadenosine 5gm 209 25gm

14284 2’-Deoxycytidine,Hydrochloride,Synthetic 1gm 210 5gm 10gm 100gm

14300 2’-Deoxyguanosine 5gm 210 25gm 100gm

14304 2’-Deoxyguanosine,MonohydrateSynthetic 1gm 210 5gm 10gm 100gm 1kg

14340 DNaseI 10mg 213 20mg

14365 DNaseI,Lyophilized 5mg 72

14367 DNaseI,Solution 100units 72 500units

14375 DNA,CalfThymus 1gm 128 5gm

14377 DNA,FishSperm,SodiumSalt 100gm 128 1kg

14380 DNA,E. coli 10mg 128

14394 DNA-Cellulose,Double-Stranded 5gm 124 10gm

14395 DNA-Cellulose,Single-Stranded 5gm 124 10gm

14405 DNA,FishSperm 100gm 128 1kg

14420 2-Deoxy-D-Ribose,Ultrapure 5gm 211 25gm 100gm

14489 DextranSulfate,SodiumSalt 10gm 211 50gm 100gm

14495 Dextran,Ultrapure 100gm 211 1kg

14535 D-(+)-Dextrose,Anhydrous 5lb 211 25lb 100lb

14536 D-(+)-Dextrose,Monohydrate 10kg 212

14564 DAPI 10mg 209 50mg

14615 Diaphorase 600units 212 1,800units

14710 DiethylPyrocarbonate,Ultrapure 25ml 212 100ml

14765 DihydrostreptomycinSulfate 100gm 213

14850 Dimedone 100gm 213 1kg

14868 p-DimethylaminoBenzaldehyde 100gm 213 500gm

14872 p-DimethylaminoCinnamaldehyde 25gm 213

15335 b-NicotinamideAdenineDinucleotide 1gm 229 5gm 10gm 25gm 100gm

15345 b-NicotinamideAdenineDinucleotide-Reduced, 1gm 230 Disodium,Trihydrate 5gm 10gm 25gm

15385 Dithioerythritol,Ultrapure 5gm 213 25gm 100gm

15395 Dithiothreitol 5gm 213 25gm 100gm

15397 Dithiothreitol,Ultrapure 1gm 213 5gm 25gm 100gm

15460 Albumin,Egg 1lb 199 5lb 25lb

15475 Elastase 5mg 214 10mg 20mg 100mg

15515 Enolase 10,000units 214

15562 Ergocalciferol 5gm 214 25gm 100gm

15590 Esculin 100gm 214

15694 EthylenediaminetetraaceticAcid,(EDTA),0.5M 100ml 215 Solution,Ultrapure 500ml

15697 EthylenediaminetetraaceticAcid,(EDTA)Disodium 1kg 215 Salt,Dihydrate

15699 EthylenediaminetetraaceticAcid,(EDTA)Disodium 500gm 215 Salt,Dihydrate 5kg

15700 EthyleneDiamineTetraaceticAcid,(EDTA)Tetrasodium 1kg 215 Salt,Dihydrate,Ultrapure




15701 EthylenediaminetetraaceticAcid,(EDTA)Disodium 100gm 215 Salt,Dihydrate,Ultrapure 500gm 1kg

15703 EthyleneGlycol-O,O’-Bis(2Amino-ethyl)-N,N,N’,N’,- 10gm 215 TetraaceticAcid(EGTA),Ultrapure 50gm 100gm

15762 FetalBovineSerum(FBS) 500mI 215

15785 FlavinAdenineDinucleotide,DisodiumSalt,Hydrate 250mg 216 500mg 1gm

15880 FolicAcid 25gm 216 100gm 1kg

15929 D-Fructose-6-Phosphate,DisodiumSalt 1gm 216 5gm

15995 D-(+)-Galactose 500gm 217 1kg 5kg

16021 g-Globulins,FractionII 10gm 217 25gm

16051 GentamicinSulfate 1gm 217 5gm 25gm

16146 GlucoseOxidase 50,000units 217 250,000units 1mu

16171 Glucose-6-PhosphateDehydrogenase 1,000units 217 Source:Leuconostoc mesenteroides 5,000units 10,000units

16176 Glucose-6-PhosphateDehydrogenase 1,000units 218 Source:Yeast 5,000units 10,000units

16185 D-Glucose-6-Phosphate,DisodiumSalt,Dihydrate 5gm 217 10gm

16215 L-GlutamicAcid 1kg 218 5kg 25kg

16245 L-GlutamicAcid,MonosodiumSalt,Monohydrate 5kg 218

16250 Glucose-6-PhosphateGlutamateDehydrogenase 100mg 218 500mg 1gm

16285 L-Glutamine 100gm 218 250gm 500gm 1kg

16315 Glutathione-Reduced 25gm 218 100gm

16320 Glutathione-Oxidized 250mg 218 1gm

16372 Glycine 2.5kg 220

16374 Glycerol,Ultrapure 500ml 219 1L

16405 Glycine 1kg 220 5kg 25kg

16407 Glycine,Ultrapure 500gm 219 1kg 5kg

16445 Glycogen,Ultrapure 5gm 220 10gm 25gm

16505 Glycyl-Glycine 100gm 220 1kg

16785 Guanosine-5’-Diphosphate,DisodiumSalt 100mg 221 500mg 1gm

16800 Guanosine-5’-Triphosphate,DisodiumSalt 25mg 221 100mg 500mg 1gm

16880 Hemin 10gm 221 50gm

16920 Heparin,SodiumSalt 50,000units 221 100,000units 500,000units 1mu

16924 HEPES,1MSolution,pH7.3,Ultrapure 100ml 222 500ml 1L

16926 HEPES,Ultrapure 100gm 222 250gm 1kg 5kg

16928 HEPES,SodiumSalt 100gm 222 1kg

16965 Hexokinase 1,000units 222 10,000units 25,000units 50,000units

17070 L-Histidine,FreeBase 100gm 222 1kg

17080 L-Histidine,Monohydrochloride,Monohydrate 100gm 222 1kg 5kg

17525 Imidazole,Ultrapure 100gm 223 500gm 1kg 5kg

17798 IsocitrateDehydrogenase,(NADP+Specific), 150units 223 Recombinant 600units

17820 DL-Isoleucine 100gm 223 500gm 1kg

17825 L-Isoleucine 10gm 223 100gm 500gm 1kg

17886 Isopropyl-b-D-Thiogalactopyranoside(IPTG),Dioxane 1gm 223 Free,Ultrapure 5gm 25gm 100gm

17924 KanamycinSulfate 5gm 223 25gm




17950 a-KetoglutaricAcid,FreeAcid 100gm 224 500gm 1kg

17955 a-KetoglutaricAcid,DisodiumSalt,Dihydrate 5gm 224 100gm 1kg

18160 L-(+)-LacticAcid,LithiumSalt 100gm 224

18177 Lactoferrin 100mg 224 500mg 1gm

18185 D-(+)-Lactose,Monohydrate 5kg 224

18218 N-DodecylTrimethylammoniumBromide 10gm 214 50gm 250gm

18220 SodiumDodecylSulfate(SDS) 500gm 240 1kg 5kg

18245 Lecithin 500gm 224 1kg

18270 DL-Leucine 100gm 225 1kg

18280 L-Leucine 100gm 225 1kg 5kg

18413 Leupeptin,Ultrapure 5mg 225 25mg 100mg

18425 b-D-Fructose 1kg 216

18505 DL-a-LipoicAcid 25gm 225 50gm 100gm

18522 DL-a-LipoicAcid 50gm 225 100gm 1kg

18585 L-Lysine,Hydrochloride 1kg 226 5kg 10kg

18641 MagnesiumChloride,Hexahydrate,Ultrapure 500gm 226 1kg

18645 Lysozyme,Ultrapure 5gm 226 10gm 25gm 100gm

18670 MalateDehydrogenase,Lyophilized 25,000units 226 100,000units

18685 MaleicAcid 1kg 226

18695 DL-MalicAcid 1kg 226

18700 L-MalicAcid 500gm 227 1kg

18724 Maltose,Monohydrate 500gm 227 1kg

18725 D-(+)-Maltose,Monohydrate 1kg 227 5kg

18755 Mannitol 500gm 227 1kg 5kg

18775 D-(+)-Mannose 100gm 227 500gm

18886 MES,Monohydrate,Ultrapure 100gm 227 1kg

18888 MES,Anhydrous 100gm 227 250gm 1kg

18893 MES,SodiumSalt 100gm 227 250gm 1kg

19184 5-Methyl-2’-Deoxycytidine-5’-Monophosphate, 100mg 228 DisodiumSalt

19220 MethyleneBlue,Trihydrate 100gm 228 500gm

19256 MOPS,Ultrapure 100gm 228 1kg

19265 MTT,Ultrapure 500mg 228 1gm 5gm 10gm

19391 a-NaphthylPhosphate,MonosodiumSalt 25gm 229

19430 Neocuproine,Hydrochloride 10gm 229 25gm 100gm 1kg

19435 NeomycinSulfate,Ultrapure 10gm 229 25gm

19438 b-NicotinamideAdenineDinucleotide,LithiumSalt, 1gm 229 Dihydrate 5gm

19439 b-NicotinamideAdenineDinucleotide,Monosodium 5gm 230 Salt

19465 NeutralRed 25gm 229 100gm 1kg

19480 Niacinamide 1kg 229 10kg

19535 p-NitroBlueTetrazoliumChloride,Ultrapure 250mg 230 1gm 5gm 10gm

19594 4-Nitrophenyl-b-D-Glucopyranoside 1gm 231 5gm

19629 SSC,20XSolution,Ultrapure 500ml 241 1L 5L

19817 Oligop(dT)12-18SodiumSalt 5units129,232 25units 100units

19880 Pancreatin1X 200gm 232

19943 ParaformaldehydeSolution,4%inPBS 1L 232

19957 PolyethyleneGlycol400 500ml 235 1L




19959 PolyethyleneGlycol8000,Flakes,Ultrapure 500gm 235 1kg 5kg

19966 PolyethyleneGlycol8000,Powder 500gm 235 1kg 5kg

19972 PolyethyleneGlycol6000,Flakes 1kg 235 5kg

19985 PenicillinG,Potassium 1kg 232

20010 Pepsin 1gm 232 5gm 10gm 25gm

20015 Pepsin1:10,000 100gm 233 500gm 1kg

20037 PepstatinA,Ultrapure 5mg 233 25mg

20048 BacteriologicalPeptone,Ultrapure 1lb 202 5lb 25lb

20075 Phenolphthalein 500gm 234 1kg

20085 PhenolRed,SodiumSalt 10gm 234 25gm 50gm

20100 DL-Phenylalanine 100gm 234 1kg

20105 L-Phenylalanine 100gm 234 250gm 1kg 5kg

20203 PhenylmethylSulfonylFluoride,Ultrapure 5gm 234 25gm 100gm

20227 PotassiumPhosphate,Monobasic,Anhydrous, 500gm 236 Ultrapure 1kg

20228 PotassiumPhosphate,Monobasic 1kg 236 10kg

20229 SodiumPhosphate,Monobasic 1kg 240 5kg

20232 SodiumPhosphate,Dibasic,Heptahydrate,Ultrapure 500gm 240 1kg

20233 SodiumPhosphate,Monobasic,Monohydrate, 500gm 241 Ultrapure 1kg

20240Y PhosphodiesteraseI 100units 81

20274 PotassiumPhosphate,Dibasic,Ultrapure 500gm 236 1kg 2.5kg

20276 PotassiumPhosphate,Dibasic 1kg 236 10kg

20426 PIPES 100gm 208 1kg

20428 PIPES,DisodiumSalt 10gm 234 100gm 1kg

20450 PolyanetholesulfonicAcid,SodiumSalt 5gm 234 100gm

20539 Poly(dI-dC)•Poly(dI-dC)SodiumSalt 5units 235 25units 100units

20551 PolymixinBSulfate 10mu 235 100mu

20598 PotassiumChloride,Ultrapure 500gm 236 1kg 5kg

20611 PVP-K-30 1kg 237

20795 ProtamineSulfate 25gm 237 100gm

20995 PyruvicAcid,SodiumSalt 10gm 237 25gm 100gm 500gm

21026 DeoxycholicAcid,SodiumSalt 25gm 210 100gm

21060 D-(+)-Raffinose,Pentahydrate 100gm 237 500gm 1kg

21145 Riboflavin 25gm 238 100gm 500gm 1kg

21185 RNA,FreeAcid 100gm 238 1kg

21190 RNA,SodiumSalt 25gm 239 100gm 1kg

21195 RNaseA,Lyophilized 100mg 239 1gm 5gm

21220 D-(–)-Ribose 10gm 238 100gm

21245 T4RNALigase 600units 71 2,500units

21246 Rifampin 1gm 238 5gm

2141Y KlenowDNAPolymeraseI 300units 872141Z 1,500units

21486 DL-Serine(Non-Animal) 100gm 239 1kg

21610 SodiumAzide,Ultrapure 100gm 239 500gm

21616 SodiumCarbonate,Anhydrous 500gm 239 2.5kg

21618 SodiumChloride,Ultrapure 500gm 239 1kg 5kg 10kg




21653 N-LauroylSarcosine,SodiumSalt,Ultrapure 50gm 224 100gm

21655 b-Glycerophosphate,SodiumSalt,Pentahydrate 100gm 219 500gm 1kg

21660 SodiumPhosphate,DibasicAnhydrous 500gm 240 2.5kg

21665 SodiumSelenite 100gm 241 500gm

21695 Starch-Soluble,Ultrapure 1kg 242

21710 D-Sorbitol,Ultrapure 500gm 241 1kg 5kg

21755 Spermine,Tetrahydrochloride 5gm 241

21760 Spermidine,Trihydrochloride,Ultrapure 1gm 241 5gm

21865 StreptomycinSulfate,Ultrapure 10gm 242 100gm 1kg

21900 SuccinicAcid,FreeAcid 500gm 242 1kg 5kg

21910 SuccinicAcid,DisodiumSalt,Hexahydrate 500gm 242

21930 Sucrose,Crystalline 1kg 243 5lb 100lb

21931 Sucrose 500gm 243 2.5kg

21938 Sucrose,Ultrapure 1kg 243 5kg

21985 SulfanilicAcid 500gm 243

22086 TES,Ultrapure 100gm 245 500gm 1kg

22105 Tetracycline,Hydrochloride,Ultrapure 25gm 245 100gm 1kg

22124 TetramethylammoniumChloride,Ultrapure 1kg 245

22126 3,3’,5,5’-Tetramethylbenzidine 1gm 245 5gm

22170 Thiamine,Hydrochloride 100gm 245 500gm 1kg

22215 Thimerosal 25gm 246 100gm

22290 L-Threonine 100gm 246 1kg

22292 ThrombinHuman 1,000units 246

22293 Thrombin 25,000units 246 100,000units

22515 a,a-Trehalose,Dihydrate 100gm 246

22561 Tricine,Ultrapure 100gm 246 500gm 1kg

22565 Triethanolamine,Hydrochloride 250gm 246 1kg

22631 2,3,5-TriphenylTetrazoliumChloride 10gm 246 25gm 50gm 100gm 500gm 1kg

22637 Tris/HCl,1MSolution,pH7.0,Ultrapure 100ml 248 500ml 1L



22650 b-NicotinamideAdenineDinucleotidePhosphate- 25mg 230 Reduced,Tetrasodium,Tetrahydrate 100mg 250mg 500mg 1gm

22655 b-NicotinamideAdenineDinucleotidePhosphate, 100mg 230 Monosodium,Tetrahydrate 250mg 500mg 1gm 5gm

22660 2,4,6-Tripyridyl-s-Triazine(TPTZ) 5gm 247 25gm 100gm

22674 Tris,HighPurityGrade 500gm 247 1kg 5kg

22675 Tris,ReagentGrade 500gm 247 1kg

22676 Tris,Hydrochloride,Ultrapure 500gm 247 1kg 5kg

22710 Trypsin1:250 100gm 248 1kg

22720 Trypsin,Ultrapure 1gm 248 5gm 10gm

22725 Trypsin(TPCKTreated) 100mg 248 250mg

22765 L-Tryptophan 100gm 248 1kg

22885 Tyrosinase 100,000units 248 500,000units

22910 L-Tyrosine 100gm 248 1kg 5kg

23020 Uracil 100gm 249 1kg




23036 Urea 1kg 249 5kg

23075 UricAcid 100gm 249 500gm 1kg

23130 Uridine 1gm 249 10gm 100gm

23160 Uridine-5’-Triphosphate,TrisodiumSalt 100mg 249 1gm

23261 Vancomycin,Hydrochloride 100mg 250 250mg

23360 VitaminB12 1gm 250

23445 VitaminK1 250mg 250 1gm 5gm 10gm

23457 Wright’sStain 5gm 250 10gm 25gm 100gm

23465 XanthineMonosodiumSalt,Monohydrate 5gm 250 10gm 25gm 100gm

23513 XyleneCyanolFF,Ultrapure 25gm 251

23520 D-(+)-Xylose 500gm 251 1kg

23540 Yeast,Bakers 1lb 251 5lb 25lb

23545 YeastExtractPowder,LowDust 1kg 251 5kg

23546 Yeast,Brewers 5lb 251 25lb 100lb

23547 YeastExtractPowder,Ultrapure 1kg 251

23550 YeastHydrolysate,Enzymatic,Ultrapure 5lb 251 25lb 50lb 100lb

23551 Yeast-Torula 5lb 251 25lb 100lb

23660 Chloramphenicol,Ultrapure 25gm 206 100gm 1kg

25110 LeptomycinB 1µg 224

26004 Thioredoxin,Human,Recombinant 100µg 246

26200 5-Bromo-4-Chloro-3-Indolyl-a-D-Galactopyranoside, 100mg 203 Ultrapure 500mg

27-0323-01 RNaseI‘A’,Powder 100mg 79,163

27-0330-02 RNaseI 1gm 79,163

32800 Acrylamide 25gm 188 100gm 500gm 1kg

32802 Agarose-LE,Ultrapure 25gm 194 100gm 250gm 500gm 1kg

32803 Agarose-ME,Ultrapure 25gm 192 100gm 250gm 500gm

32804 Agarose-HE,Ultrapure 100gm 192 250gm 500gm

32812 BrilliantBlueG,Ultrapure 25gm 203 100gm

32813 EthidiumBromide,Ultrapure 1gm 214 5gm 25gm

32816 LithiumDodecylSulfate,Ultrapure 25gm 225 100gm

32819 PonceauS,SodiumSalt,Ultrapure 50gm 236

32821 Agarose-UltraLowMelt,Ultrapure 10gm 197 25gm

32823 Starch,Hydrolyzed,Ultrapure 100gm 242 1kg

32826 BrilliantBlueR,Ultrapure 25gm 203 100gm

32829 Agarose-LowMeltSeparation≤1000bp,Genetic 25gm 197 PerformanceCertified,Ultrapure 100gm

32830 Agarose-LowMeltSeparation≥1000bp,Genetic 25gm 196 PerformanceCertified,Ultrapure 100gm

33230 4-Methylumbelliferyl-a-L-Iduronide,SodiumSalt 5mg 228

34007 GuanidineHydrochloride 1kg 221 5kg

34128 4-Nitrophenyl-N-Acetyl-b-D-Galactosaminide 100mg 230 1gm

34131 4-Nitrophenyl-a-L-Fucopyranoside 50mg 231 250mg 1gm

34133 4-Nitrophenyl-a-D-Galactopyranoside 500mg 231 1gm 5gm

34136 4-Nitrophenyl-b-D-Glucuronide 100mg 231 250mg 1gm

34138 4-Nitrophenyl-a-D-Maltoside 50mg 231 100mg

34140 4-Nitrophenyl-b-D-Mannopyranoside 100mg 231 500mg

34142 4-Nitrophenyl-b-D-Xylopyranoside 500mg 231 1gm




70001Z T7RNAPolymerase,Highconcentration, 30,000units 92,161 >80units/µl

70005Y T4DNALigase,Standardconcentration,1unit/µl 100units 7070005X 500units70005 2,000units

70010Y DNAPolymeraseI 500units 87

70017Z T7DNAPolymerase 250units 8870017 1,000units

70020Y E. coliDNALigase 200units 7070020Z 1,000units

70023Y ExonucleaseIII 5,000units 7770023Z 25,000units

70025Y T7Gene6Exonuclease 2,000units 7870025Z 10,000units

70028Y RecAProtein 200µg 6570028Z 1,000µg

70029Y T4Gene32Protein,Standardconcentration,5µg/µl 100µg 6670029Z 500µg

70031Y T4PolynucleotideKinase(PNK) 500units 6970031Z 1,000units70031X 2,500units

70032Y Single-StrandedDNABindingProtein(SSB) 100µg 6570032Z 500µg

70034Y CalfIntestinalAlkalinePhosphatase(CIAP) 1,500units 82

70041Y AMVReverseTranscriptase 200units 90,16270041Z 1,000units

70042X T4DNALigase,Highconcentration,10units/µl 500units 7070042 2,000units

70047Y T7RNAPolymerase,Standardconcentration, 6,000units 92,161 20units/µl

70051Y T3RNAPolymerase,Standardconcentration, 1,000units 92,16070051Z 20units/µl 5,000units

70052 TthDNAPolymerase 200units 89 1,000units

70054Y RNaseH 250units 80,16370054Z 1,000units

70057Y Exonuclease-FreeKlenow 125units 8670057Z 750units70057 2,500units

70065 7-deaza-2’-Deoxyguanosine-5’-Triphosphate, 2µmol 60,210 LithiumSalt,10mMSolution

70073Z ExonucleaseI,Standardconcentration,10units/µl 2,500units 7670073X 5,000units

70076 DNA,FishSperm,Activated 500µg 128 2mg

70082Y ExonucleaseVII 200units 7770082Z 1,000units

70140 SequenaseQuickDenaturePlasmidDNA 100reactions 177 SequencingKit

70170 SequenasePCRProductSequencingKit 100reactions 176

70175 SequenaseVersion2.0DNAPolymerasewith 325units 85 Pyrophosphatase

70194Y RNaseA,MolecularBiologyGrade,Solution 1mg 80,16370194Z 5mg

70196Y MicrococcalNuclease 15,000units 81

70204 T3RNAPolymerase,Highconcentration, 1,000units 92,160 200units/µl 5,000units

70440 T4RNALigase2 200units 71 1,000units

70454 TBEBuffer,10XReady-MixedPowder,Ultrapure 6x200ml 243

70468 Denhardt’sSolution,50X,Ultrapure 50ml 209

70704 M13mp18,Single-Stranded,0.2µg/ul 50µl 125 50µg

70724 StopSolution,Ultrapure 1,200µl 242

70726 Dithiothreitol(DTT),0.1MSolution,Ultrapure 150µl 213 1ml 5ml

70770 SequenaseVersion2.0DNASequencingKit 100reactions 175

70775Y SequenaseVersion2.0DNAPolymerase 200units 88,17470775Z 1,000units

70783 Water,RNase-Free,DEPCTreated,Ultrapure 25ml 250 10x1ml 100ml 500ml 1L

70796 DextranSulfate,SodiumSalt,Ultrapure 10gm 211 50gm

70995 PCRProductPre-SequencingKit 100reactions 5270996 500reactions70997 2,000reactions

71155 HotStart-ITFideliTaqDNAPolymerase 50units 17,25 250units 1,000units 5,000units

71156 HotStart-ITFideliTaqPCRMasterMix(2X) 25reactions 18 100reactions 500reactions

71160 TaqDNAPolymerase 50units 11 250units 1,000units 5,000units

71161 TaqPCRKit 100reactions(100µl/rctn) 12

71162 TaqPCRMasterMix(2X) 100reactions 12

71170 VeriQuestTaqDNAPolymerase 50units 26 250units 1,000units 5,000units

71180 FideliTaqDNAPolymerase 50units 15 250units 1,000units 5x250units 5,000units

71182 FideliTaqPCRMasterMix(2X) 100reactions 16




71185 FideliTaqRT-PCRMasterMix(2X) 100reactions 46

71190 ROXPassiveReferenceDye 50units 13

71191 RubyTaqPCRMasterMix(2X) 100reactions 14 500reactions

71194 HotStart-ITBindingProtein 400μg 21

71195 HotStart-ITTaqDNAPolymerase 50units 19 250units 1,000units 5x250units 5,000units

71196 HotStart-ITTaqPCRMasterMix(2X) 25reactions 20 100reactions 500reactions

71199 MagniTaqMultiplexPCRMasterMix(2X) 25reactions 22 100reactions

71200 T3PromoterPrimer 250pmol 178

71547 Oligop(dT)Cellulose,Ultrapure 1gm 125

71570 RNaseInhibitor(HumanPlacenta) 5,000units 94,164

71571 RNaseInhibitor(Recombinant) 5,000units 94,164

71600 MineralOil,Ultrapure 10ml 228 25ml 1L

71706 M13mp18DNA,Single-Stranded,1.0µg/µl 50µg 125 200µg

71786 Water,Nuclease-Free,Ultrapure 10x1ml 250 100ml 500ml 1L 5L

71960 Uracil-DNAGlycosylase,E. coli 400units 67 2,000units

72020 Exonuclease-FreeKlenow,TrisBuffer 750units 86 2,500units

72033 TerminalDeoxynucleotidylTransferase, 500units 93 Recombinant(rTdT) 2,500units

72073 ExonucleaseI,Highconcentration,20units/µl 5,000units 76

74015 TAEBuffer,50XSolution,Ultrapure 100ml 243

74029Y T4Gene32Protein,Highconcentration,≥10µg/µl 300µg 6674029Z 1,000µg

74225Y Poly(A)Polymerase,Yeast 10,000units 91,15974225Z 25,000units

75592 SuperSAPShrimpAlkalinePhosphatase 50reactions 84

75600 VeriQuestSYBRGreenqPCRMaster 40reactions(1ml) 27 Mix(2X) 200reactions(5ml) 400reactions(2x5ml) 1,000reactions(5x5ml) 2,000reactions(10x5ml)

75650 VeriQuestProbeqPCRMasterMix(2X) 40reactions(1ml) 33 200reactions(5ml) 400reactions(2x5ml) 1,000reactions(5x5ml) 2,000reactions(10x5ml)

75660 VeriQuestProbeqPCRMasterMix, 40reactions(1ml) 35 NoReferenceDye(2X) 200reactions(5ml) 400reactions(2x5ml) 1,000reactions(5x5ml) 2,000reactions(10x5ml)

75665 VeriQuestSYBRGreenqPCRMaster 40reactions(1ml) 29 MixwithFluorescein(2X) 200reactions(5ml) 400reactions(2x5ml) 1,000reactions(5x5ml) 2,000reactions(10x5ml)

75675 VeriQuestFastSYBRGreenqPCR 100reactions(1ml) 31 MasterMixwithFluorescein(2X) 500reactions(5ml) 1,000reactions(2x5ml) 2,500reactions(5x5ml) 5,000reactions(10x5ml)

75680 VeriQuestFastProbeqPCRMasterMix(2X) 100reactions(1ml) 36 500reactions(5ml) 1,000reactions(2x5ml) 2,500reactions(5x5ml) 5,000reactions(10x5ml)

75685 VeriQuestFastProbeqPCRMaster 100reactions(1ml) 37 Mix,NoReferenceDye(2X) 500reactions(5ml) 1,000reactions(2x5ml) 2,500reactions(5x5ml) 5,000reactions(10x5ml)

75690 VeriQuestFastSYBRGreenqPCR 100reactions(1ml) 30 MasterMix(2X) 500reactions(5ml) 1,000reactions(2x5ml) 2,500reactions(5x5ml) 5,000reactions(10x5ml)

75700 VeriQuestProbeOne-StepqRT-PCR 40reactions(1ml) 42 MasterMix(2X) 200reactions(5ml)

75705 VeriQuestSYBRGreenOne-StepqRT-PCR 40reactions(1ml) 40 MasterMix(2X) 200reactions(5ml)

75710 VeriQuestProbeOne-StepqRT-PCR 40reactions(1ml) 43 MasterMix,NoReferenceDye(2X) 200reactions(5ml)

75715 VeriQuestSYBRGreenOne-StepqRT-PCR 40reactions(1ml) 41 MasterMixwithFluorescein(2X) 200reactions(5ml)

75760 HotStart-ITSYBRGreenqPCRMasterMix 100reactions 32 withUDG(2X) 500reactions

75762 HotStart-ITSYBRGreenqPCRMasterMix(2X) 100reactions 32 500reactions

75764 HotStart-ITProbeqPCRMasterMixwith 100reactions 38 UDG(2X) 500reactions

75766 HotStart-ITProbeqPCRMasterMix(2X) 100reactions 38 500reactions

75767 FluoresceinPassiveReferenceDye 500μl 53

75768 RT-PCRMasterMix(2X) 500μl 53

75770 HotStart-ITSYBRGreenOne-StepqRT-PCRMaster 100reactions 44 MixKit 500reactions

75772 HotStart-ITProbeOne-StepqRT-PCRMaster 100reactions 44 MixKit 500reactions

75780 First-StrandcDNASynthesisKitfor 50reactions(20µl) 39 Real-TimePCR

75790 TEBuffer,1XSolution,pH7.0 100ml 244 500ml




75791 TEBuffer,1XSolution,pH7.0,LowEDTA 100ml 244 500ml

75792 TEBuffer,1XSolution,pH8.0 100ml 244 500ml

75793 TEBuffer,1XSolution,pH8.0,LowEDTA 100ml 244 500ml

75816 EthidiumBromideDrops,Ultrapure 5ml 214

75817 Agarose-Separation≥500bp,GeneticPerformance 25gm 193 Certified,Ultrapure 100gm 250gm 500gm

75818 GuanidineThiocyanate,Ultrapure 100gm 221 250gm 1kg

75819 SodiumDodecylSulfate(SDS),Ultrapure 100gm 240 1kg

75820 Acrylamide,Ultrapure 100gm 188 500gm 1kg

75821 N,N’-Methylene-bis-Acrylamide,Ultrapure 25gm 228 50gm 100gm 1kg

75822 CesiumChloride,Ultrapure 5gm 206 25gm 100gm 500gm 1kg

75823 GuanidineHydrochloride,Ultrapure 100gm 220 500gm

75824 Taurine,Ultrapure 1kg 243

75825 Tris,Ultrapure 500gm 247 1kg 5kg 10kg

75826 Urea,Ultrapure 1kg 249 5kg

75827 GlycerolTolerantGel(GTG)Buffer,20XSolution, 1L 219 Ultrapure

75828 Formamide,Ultrapure 500gm 216 1kg

75829 Phenol,pH8.0,Equilibrated,Ultrapure 100ml 233 400ml

75830 Phenol,Ultrapure 500gm 233

75831 Phenol:Chloroform:IsoamylAlcohol(25:24:1), 100ml 233 Ultrapure 400ml

75832 SodiumDodecylSulfate(SDS),20%Solution,Ultrapure 100ml 240 500ml

75834 TEBuffer,50XSolution,Ultrapure 100ml 244 500ml

75848 RapidGel40%LiquidAcrylamideStockSolution, 500ml 237 Ultrapure

75851 LBAgar,Ready-MadePowder 250gm 224 1kg

75852 LBBroth,Ready-MadePowder 250gm 224 1kg

75853 LuriaAgar,Ready-MadePowder 250gm 225 1kg

75854 LuriaBroth,Ready-MadePowder 250gm 225 1kg

75855 NZCYMBroth,Ready-MadePowder 1kg 232

75856 TerrificBroth,Ready-MadePowder 250gm 245 1kg

75861 RapidGel-XL6%LiquidAcrylamideDNASequencing 500ml 238 Gel,TBEFormulation,Ultrapure

75864 2XYTBroth 1kg 251

75888 SodiumChloride(NaCl),5MSolution,Ultrapure 100ml 239 500ml 1L

75889 PBS,10XSolution,pH7.4,Ultrapure 100ml 232 1L 5L

75890 SSPE,20XSolution,Ultrapure 1L 241 5L

75891 TBEBuffer,5XSolution,Ultrapure 1L 243 5L

75892 TBS,20XSolution,pH7.4,Ultrapure 100ml 244 1L 5L

75893 TEBuffer,1XSolution,Ultrapure 10x1ml 244 100ml 500ml

75894 Tris-Glycine(TG)Buffer,10XSolution,Ultrapure 1L 247 5L

75895 Tris-Glycine-SDS(TGS)Buffer,10XSolution,Ultrapure 1L 247 5L

75896 PotassiumChloride(KCl),2MSolution,Ultrapure 100ml 236

75897 SodiumAcetate(NaOAc),3MSolution,pH5.5, 100ml 239 Ultrapure 500ml

75901 AmmoniumAcetate(NH4OAc),5MSolution,Ultrapure 100ml 199 500ml

75902 LithiumChloride(LiCl),7.5MSolution,Ultrapure 100ml 225

75904 TAEBuffer,10XSolution,Ultrapure 1L 243 5L

76225 ProteinaseKSolution 100mg 137

76230Y ProteinaseK 100mg137,23776230Z 500mg76230 1gm(2x500mg)

76320 N,N,N’,N’-Tetramethylethylenediamine,Ultrapure 100ml 245 100gm 500gm

76322 AmmoniumPersulfate(APS),Ultrapure 10gm 200 100gm 1kg

76324 BoricAcid,Ultrapure 500gm 202 1kg




76410 LowMolecularWeightMarker,10-100nt 100µl132,165

76455 Poly(A)Tail-LengthAssayKit 1kit 158

76632 pUC19DNA,0.2µg/µl 10µg 129 50µg 200µg

76634 pUC19DNA,1.0µg/µl 50µg 129 200µg

76710 PCRMarkers,50-2000bp 250µl 130

76712 DNALadder,100bp 500µl 130

76714 DNALadder,1kbPlus 500µl 131

76715 DNALoadingBuffer(OXG),6X 1ml 130 5ml

76720 DNALoadingBuffer(BXF),6X 1ml 131

76740 ProteinMarkers,10-225kDa 500µl 136

77100 2’-Deoxynucleoside-5’-Triphosphates,SodiumSalt, 1pk 59,211 100mMSolutions,SetsofFour,Ultrapure 4x25µmol(250µl)eachdNTPperpack

77102 2’-Deoxyadenosine-5’-Triphosphate,Sodium 25µmol(250µl) 59,210 Salt,100mMSolution,Ultrapure 100µmol(1ml) 500µmol(5ml)

77104 2’-Deoxycytidine-5’-Triphosphate,SodiumSalt, 25µmol(250µl) 59,210 100mMSolution,Ultrapure 100µmol(1ml) 500µmol(5ml)

77106 2’-Deoxyguanosine-5’-Triphosphate,Sodium 25µmol(250µl) 59,210 Salt,100mMSolution,Ultrapure 100µmol(1ml) 500µmol(5ml)

77108 2’-Deoxythymidine-5’-Triphosphate,Sodium 25µmol(250µl) 59,211 Salt,100mMSolution,Ultrapure 100µmol(1ml) 500µmol(5ml)

77110 2’,3’-Dideoxyadenosine-5’-Triphosphate, 0.5µmol(50µl) 60,212 Ultrapure,10mMaqueoussolution,pH7.5 1µmol(100µl)

77112 2’,3’-Dideoxycytidine-5’-Triphosphate, 0.5µmol(50µl) 60,212 Ultrapure,10mMaqueoussolution,pH7.5 1µmol(100µl)

77116 2’,3’-Dideoxythymidine-5’-Triphosphate, 0.5µmol(50µl) 60,212 Ultrapure,10mMaqueoussolution,pH7.5 1µmol(100µl)

77119 PCRNucleotideMix,25mMSolution,Ultrapure 500µl 60,232

77125 2’,3’-Dideoxynucleoside-5’-Triphosphates,10mM 1pk 60,212 Solutions,SetsofFour,Ultrapure,4x1.0µmol(100µl) eachddNTPperpack

77126 2’,3’-Dideoxynucleoside-5’-Triphosphates,10mM 1pk 60,212 Solutions,SetsofFour,Ultrapure,4x0.5µmol(50µl) eachddNTPperpack

77128 2’-Deoxynucleoside-5’-Triphosphates,SodiumSalt, 1pk 59,211 100mMSolutions,SetsofFour,Ultrapure 4x500µmol(5ml)eachdNTPperpack

77206 2’-Deoxyuridine-5’-Triphosphate,SodiumSalt, 25µmol(250µl) 59,211 100mMSolution,Ultrapure 100µmol(1ml) 500µmol(5ml)

77212 PCRNucleotideMix,10mMSolution,Ultrapure 500µl 60,232 2x500µl

77241 Adenosine-5’-Triphosphate,100mMSolution, 25µmol(250µl) 61,189 Ultrapure

77243 Guanosine-5’-Triphosphate,SodiumSalt, 25µmol(250µl) 61,221 100mMSolution,Ultrapure

77245 Nucleoside-5’-Triphosphates(ATP,CTP,GTP,UTP), 1pk 61,231 SetofFour,100mMSolutions,Ultrapure 4x25µmol(250µl)eachNTPperpack

77328 2’-Deoxynucleoside-5’-Triphosphates,SodiumSalt, 1pk 59,211 100mMSolutions,SetsofFour,Ultrapure 4x100µmol(1ml)eachdNTPperpack

77330 PCRNucleotideMixwithdUTP,10mMSolution, 500µl 60,232 Ultrapure

77335 2’,3’-Dideoxynucleoside-5’-Triphosphates,100mM 1pk 60,212 Solutions,SetofFour,Ultrapure,4x4µmol(40µl) eachddNTPperpack

77405 Oligo(dT)12-18Primer,MW≅4500 100µl(2.5nmol)129,231

77500 TBSwithTween(TBST),20XSolution,pH7.4, 1L 244 Ultrapure 5L

77504 SodiumDodecylSulfate(SDS),10%Solution, 100ml 240 Ultrapure 500ml 1L

77507 Betaine,5MSolution,Ultrapure 1.5ml 202 5x1.5ml 10ml

77523 RapidRunAgaroseBuffer,20XSolution,Ultrapure 1L 238 5L

77524 RapidRunLoadingDye,Ultrapure 1ml 238 5ml

77531 TCEP-HCI,0.5MSolution,pH6.6±0.1 5ml 244 50ml

77534 Glycogen,20mg/ml,Ultrapure 1ml 220 5x1ml

78020Y RNaseA,Lyophilized 100mg 79,163

78040Y E. coliRNAPolymeraseHoloenzyme 100units 91,159

78250 ExoSAP-ITPCRProductCleanup 20reactions 49,14878200 100reactions78201 500reactions78202 2,000reactions78205 5,000reactions

78260 SBECleanupReagent 864µl 52

78300 T4EndonucleaseVII 50,000units 75

78303Y TopoisomeraseII,Alpha 200units 9578303Z 500units

78306 M-MLVReverseTranscriptase 25,000units 90,162 100,000units

78310 Uracil-DNAGlycosylase,Heat-Labile 100units 67

78314 ShrimpDNase,Recombinant 100units 74 500units

78331 10XDNaseIBuffer 1ml 74

78334X OptiKinase 500units 6878334Y 1,000units78334Z 2,500units

78350 One-StepRT-PCRKit 50reactions 47




78355 Two-StepRT-PCRKit 50RTrctnsand 48 100PCRrctns

78370 RubyTaqDNAPolymerase 100reactions 45 250units 1,000units 5,000units

78390 ShrimpAlkalinePhosphatase(SAP) 500units 83 1,000units 5,000units

78395 HTExoSAP-ITHigh-ThroughputPCR 480rctnsx8-tubestrip 50,149 ProductCleanup 5,760rctnsx1plate (12x8-tubestrips) 23,040rctns-4plates 1,000rctns(2ml) 5,000rctns(10ml)

78400 Ligate-ITRapidLigationKit 25reactions 12578410 100reactions

78411 rDNaseI,RNase-Free 1,000units 73 2,500units

78430 SequenaseRandomPrimerLabelingKit 30reactions 127

78450 Pyrophosphatase,Inorganic(Recombinant)(rPPase) 5units 85 50units

78454 HumanApurinic/ApyrimidinicEndonuclease1 1,000units 75 (APE1) 5,000units

78480 Change-ITMultipleMutationSiteDirected 20reactions 126 MutagenesisKit

78500 ThermoSequenaseCycleSequencingKit 100reactions 172

78641 MagnesiumChloride(MgCl2),1MSolution, 10x1ml 226 Ultrapure 100ml

78706 PrepEaseMidiPlasmidKit 100preps 140

78711 PrepEaseMaxiPlasmidKit 50preps 140

78720 PrepEaseGigaPlasmidKit 5preps 140

78722 PrepEaseBACPurificationKit 10preps 145

78730 PrepEaseEndotoxin-FreeMaxiPlasmidKit 10preps 142

78737 PrepEaseMiniSpinPlasmidKit 250preps 141

78742 PrepEaseQuickMiniSpinPlasmidKit 250preps 141

78745 PrepEasePlasmid8-wellStripKit 12strips 143

78751 PrepEasePlasmid96-wellPlateKit 4x96-wellplates 144

78753 PrepEasePlasmid96-wellPlateCoreKit 24x96-wellplates 144

78756 PrepEaseGelExtractionKits 50preps 14678757 250preps

78758 PrepEaseDNACleanupKits 50preps 14678759 250preps

78761 PrepEasePCRPurification96-WellPlateKits 10x96-wellplates 14878762 (Ultrafiltration) 50x96-wellplates

78766 PrepEaseRNASpinKits 50preps 15078767 250preps

78776 PrepEaseSequencingDyeCleanupKits 50preps 15278777 250preps

78780 PrepEaseVacuumManifold 1ea 143

78784 PrepEase8-wellStripStarterKitforVacuum 1pack 143

78791 PrepEaseHistidine-TaggedProteinPurificationMini 50preps 153 Kit-HighSpecificity

78793 PrepEaseHistidine-TaggedProteinPurificationMidi 50preps 153 Kit-HighSpecificity

78795 PrepEaseHistidine-TaggedProteinPurificationMaxi 25preps 153 Kit-HighSpecificity

78796 PrepEaseHistidine-TaggedHighSpecificity 30gm 155 PurificationResin

78801 PrepEaseHistidine-TaggedProteinPurificationMini 50preps 154 Kit-HighYield

78803 PrepEaseHistidine-TaggedProteinPurificationMidi 50preps 154 Kit-HighYield

78805 PrepEaseHistidine-TaggedProteinPurificationMaxi 25preps 154 Kit-HighYield

78806 PrepEaseHistidine-TaggedHighYieldPurification 30gm 155 Resin 120gm

78820 PrepEaseProteinPurificationGlutathioneAgarose4B 10ml 155 100ml

78850 PrepEaseGenomicDNAIsolationKits 50preps 14778855 250preps

78871 PrepEaseRNA/ProteinSpinKit 50preps 151

78878 PrepEasemRNAMiniSpinKit 12preps 150

79010 Biotin-11-ψTPAnalog(RNALabeling 250nmol(10mM) 57 Reagent,RLR) 1,000nmol(10mM)

79015 Biotin-11-dXTPAnalog(DNALabeling) 250nmol(10mM) 57 Reagent,DLR 1,000nmol(10mM)

79220 PrepEaseYeastPlasmidIsolationKit 50preps 145

79260 ThermoSequenaseDyePrimerManualCycle 50reactions 173 SequencingKit

798001KT Prep2SeqMultiplexOligoAdaptersforIllumina® Set1124,171 platform,(Containsadaptersequences1-12)

798002KT Prep2SeqMultiplexOligoAdaptersforIllumina® Set2124,171 platform,(Containsadaptersequences13-27)

79900 Prep2SeqDNALibraryPrepKitfor 10reactions123,170 Illumina®platform 20reactions

AY1000 EMSAKitwithoutProbeSet kit 103

AY1XXX* EMSAKit-*Seepage104forfulllisting kit 103

AY1XXXP* EMSATFProbeSets-*Seepage104forfulllisting ea 103

AY2002 NuclearExtractionKit 20rxnfromcellculture; 102 10rxnfromwholetissue

AY9999 EMSAComboKit(anythreeprobes) kit 103

BC1009 AFPELISAKit kit 134

BC1013 CA125ELISAKit kit 134

BC1015 CA15-3ELISAKit kit 134

BC1017 CA19-9ELISAKit kit 134

CI0002 CancerCellIsolationKit(2) 2assays 101






DX0001 DeliverXsiRNATransfectionEvaluationKit 0.12ml 98

DX0002 DeliverXsiRNATransfectionKit 0.4ml 98

DX0003 DeliverXsiRNATransfectionKit 1.0ml 98

DX0004 DeliverXsiRNATransfectionKit 4x1.0ml 98

DX0051 DeliverXPlussiRNATransfectionEvaluationKit 0.12ml 98

DX0052 DeliverXPlussiRNATransfectionKit 0.4ml 98

DX0053 DeliverXPlussiRNATransfectionKit 1.0ml 98

DX0054 DeliverXPlussiRNATransfectionKit 4x1.0ml 98

DX0100 FAM-labeledsiRNAControl 0.12ml 98

DX0101 HumanGAPDHsiRNAControl 0.06ml 98

DX0500 DeliverXsiRNABuffer-1 4ml 98






DX0551 DeliverXPlussiRNABuffer-1 4ml 98






DX1001 DeliverXPeptideTransfectionEvaluationKit 0.6ml 99

DX1002 DeliverXPeptideTransfectionKit 3.0ml 99

DX1100 DeliverXTAMRA-labeledPeptideControl 75µl 99

DX1101 DeliverXPre-formedPeptideTransfectionComplexControl 1.5ml 99

DX1500 DeliverXPeptideBuffer-1 4ml 99




EK0500 HIFColdControlProbe 25μl 110

EK0501 HIFControlCos1NuclearExtract 20μl 110

EK0502 EREColdProbe 25μl 110

EK0503 ERControlMCF7NuclearExtract 20μl 110

EK0504 AP-1ColdProbe 25μl 110

EK0505 ControlHEK292PMANuclearExtract 20μl 110

EK0506 p53ColdControlProbe 25μl 110

EK0508 GRColdControlProbe 25μl 110

EK0509 GRControlHela(DEX)NuclearExtract 20μl 110

EK0510 STAT-1ColdControlProbe 25μl 110

EK0511 STAT-1ControlMCF7(IFNg) 20μl 110

EK0512 MEF-2ColdControlProbe 25μl 110

EK0513 MEF-2ControlNuclearExtract 20μl 110

EK0514 SP-1ColdProbe 25μl 110

EK0515 SP-1ControlRecombinantProtein 20μl 110

EK0516 PPARColdControlProbe 25μl 110

EK0518 PPARaColdControlProbe 25μl 110

EK0519 PPARaControlCos1NuclearExtract 20μl 110

EK0520 GATA-1ColdProbe 25μl 110

EK0521 GATA-1ControlRecombinantProtein 20μl 110

EK0522 NFkBColdControlProbe 25μl 110

EK0523 NFkBControlNuclearExtract 20μl 110

EK1020 HIF-1aELISAKit kit 110

EK1021 HIF-1aELISAKitwithNuclearExtractionKit kit 110

EK1030 ERaELISAKit kit 110

EK1031 ERaELISAKitwithNuclearExtractionKit kit 110

EK1040 AP-1ELISAKit kit 110

EK1041 AP-1ELISAKitwithNuclearExtractionKit kit 110

EK1050 p53ELISAKit kit 110

EK1051 p53ELISAKitwithNuclearExtractionKit kit 112

EK1060 GRELISAKit kit 110

EK1061 GRELISAKitwithNuclearExtractionKit kit 110

EK1070 STAT-1ELISAKit kit 110

EK1071 STAT-1ELISAKitwithNuclearExtractionKit kit 110

EK1080 MEF-2ELISAKit kit 110

EK1081 MEF-2ELISAKitwithNuclearExtractionKit kit 110

EK1090 SP-1ELISAKit kit 110

EK1091 SP-1ELISAKitwithNuclearExtractionKit kit 110

EK1100 GATA-1ELISAKit kit 110

EK1101 GATA-1ELISAKitwithNuclearExtractionKit kit 110

EK1110 NFkBp50ELISAKit kit 110

EK1111 NFkBp50ELISAKitwithNuclearExtractionKit kit 110



EK1310 PPARaELISAKit kit 110

EK1311 PPARaELISAKitwithNuclearExtractionKit kit 110




EK1400 FKHRELISAKit kit 110

EK1401 FKHRELISAKitwithNuclearExtractionKit kit 110

EK1500 SMAD-4ELISAKit kit 110

EK1501 SMAD-4ELISAKitwithNuclearExtractionKit kit 110

EK1600 SMAD-2ELISAKit kit 110

EK1601 SMAD-2ELISAKitwithNuclearExtractionKit kit 110

EK1800 HNF-3bELISAKit kit 110

EK1801 HNF-3bELISAKitwithNuclearExtractionKit kit 110

EK2000 E2F-1ELISAKit kit 110

EK2001 E2F-1ELISAKitwithNuclearExtractionKit kit 110



EK4100 NFkBc-RelELISAKit kit 110

EK4101 NFkBc-RelELISAKitwithNuclearExtractionKit kit 110

EK5100 NFkBRel-bELISAKit kit 110

EK5101 NFkBRel-bELISAKitwithNuclearExtractionKit kit 110

LR0000 TA-luc(EmptyControl)LuciferaseReporterVector 20µg 112

LR0002 AP1(1)LuciferaseReporterVector 10µg 112

LR0003 AP1(2)LuciferaseReporterVector 10µg 112

LR0005 AP3LuciferaseReporterVector 10µg 112

LR0007 ARLuciferaseReporterVector 10µg 112

LR0016 E2F(1)LuciferaseReporterVector 10µg 112

LR0017 E2F(2)LuciferaseReporterVector 10µg 112

LR0019 ELK1LuciferaseReporterVector 10µg 112

LR0020 ERLuciferaseReporterVector 10µg 112

LR0025 GASLuciferaseReporterVector 10µg 112

LR0026 GAS/ISRELuciferaseReporterVector 10µg 112

LR0027 GATALuciferaseReporterVector 10µg 112

LR0028 GATA1LuciferaseReporterVector 10µg 112

LR0029 GATA1/GATA2LuciferaseReporterVector 10µg 112




LR0033 GRLuciferaseReporterVector 10µg 112

LR0038 HSELuciferaseReporterVector 10µg 112

LR0039 IRF1LuciferaseReporterVector 10µg 112

LR0040 ISRELuciferaseReporterVector 10µg 112

LR0043 MEF1LuciferaseReporterVector 10µg 112



LR0050 NF-ATLuciferaseReporterVector 10µg 112

LR0051 NFkB(1)LuciferaseReporterVector 10µg 112

LR0052 NFkB(2)LuciferaseReporterVector 10µg 112

LR0057 p53LuciferaseReporterVector 10µg 112

LR0067 PRLuciferaseReporterVector 10µg 112

LR0068 RARLuciferaseReporterVector 10µg 112

LR0070 RXRLuciferaseReporterVector 10µg 112

LR0071 Sp1LuciferaseReporterVector 10µg 112

LR0072 SRELuciferaseReporterVector 10µg 112

LR0073 SRFLuciferaseReporterVector 10µg 112

LR0075 STAT1p84/p91LuciferaseReporterVector 10µg 112

LR0077 STAT3(1)LuciferaseReporterVector 10µg 112

LR0079 STAT4LuciferaseReporterVector 10µg 112

LR0087 VDRLuciferaseReporterVector 10µg 112

LR0090 YY1LuciferaseReporterVector 10µg 112

LR0093 CRE(1)LuciferaseReporterVector 10µg 112

LR0094 E2F1(3)LuciferaseReporterVector 10µg 112

LR0114 RXR(2)LuciferaseReporterVector 10µg 112

LR0115 Smad3/Smad4LuciferaseReporterVector 10µg 112

LR0116 SmadLuciferaseReporterVector 10µg 112

LR0117 SP1(2)LuciferaseReporterVector 10µg 112

LR0127 STAT1(2)LuciferaseReporterVector 10µg 112

LR0128 HIF1LuciferaseReporterVector 10µg 112

LR1001 ABL1GenePromoterReporterVector 10µg 111

LR1002 BRCA1GenePromoterReporterVector 10µg 111

LR1003 CGPR(CALCR)GenePromoterReporterVector 10µg 111

LR1004 GAGE1GenePromoterReporterVector 10µg 111

LR1005 IRF7GenePromoterReporterVector 10µg 111

LR1006 POMCGenePromoterReporterVector 10µg 111

LR1007 ATF2GenePromoterReporterVector 10µg 111

LR1008 STAT1GenePromoterReporterVector 10µg 111

LR1009 14-3-3-sigma(SFN)GenePromoterReporterVector 10µg 111

LR1010 CIITAGenePromoterReporterVector 10µg 111

LR1011 IFNgGenePromoterReporterVector 10µg 111

LR1012 IGRPGenePromoterReporterVector 10µg 111

LR1013 IL1aGenePromoterReporterVector 10µg 111

LR1014 IL2GenePromoterReporterVector 10µg 111

LR1015 TNFaGenePromoterReporterVector 10µg 111




LR1016 SURVIVINGenePromoterReporterVector 10µg 111

LR1017 ERaGenePromoterReporterVector 10µg 111

LR1018 RB1GenePromoterReporterVector 10µg 111

LR1019 APCGenePromoterReporterVector 10µg 111

LR1020 CFTRGenePromoterReporterVector 10µg 111

LR1021 G6PDGenePromoterReporterVector 10µg 111

LR1022 hMLH1GenePromoterReporterVector 10µg 111

LR1023 IL4GenePromoterReporterVector 10µg 111

LR1024 JUNBGenePromoterReporterVector 10µg 111

LR1025 MyoDGenePromoterReporterVector 10µg 111

LR1026 VHLGenePromoterReporterVector 10µg 111

LR1027 v-mycGenePromoterReporterVector 10µg 111

LR1028 MDR1GenePromoterReporterVector 10µg 111

LR1029 COX2(PTGS2)GenePromoterReporterVector 10µg 111

LR1030 HOXA2GenePromoterReporterVector 10µg 111

LR1031 HOXA5GenePromoterReporterVector 10µg 111

LR1032 FHITGenePromoterReporterVector 10µg 111

LR1033 THBS2GenePromoterReporterVector 10µg 111

LR1034 TIMP3GenePromoterReporterVector 10µg 111

MA1210 Protein/DNAArrayI kit 116

MA1211 Protein/DNAArrayII kit 116

MA1212 Protein/DNAArrayIII kit 116

MA1213 Protein/DNAArrayIV kit 116

MA1214 Protein/DNAArrayV kit 116

MA1215 ComboProtein/DNAArray kit 116

MA1310 Protein/DNAArrayIRefillKit kit 116

MA1311 Protein/DNAArrayIIRefillKit kit 116

MA1312 Protein/DNAArrayIIIRefillKit kit 116

MA1313 Protein/DNAArrayIVRefillKit kit 116

MA1314 Protein/DNAArrayVRefillKit kit 116

MA1315 ComboProtein/DNARefillKit kit 116

MA1500 cAMP/CalciumProtein/DNAArray kit 115

MA1505 CellGrowthProtein/DNAArray kit 115

MA1510 NuclearReceptorProtein/DNAArray kit 115

MA1600 cAMP/CalciumProtein/DNAArrayRefill kit 115

MA1605 CellGrowthProtein/DNAArrayRefill kit 115

MA1610 NuclearReceptorProtein/DNAArrayRefill kit 115

MA5010 TF-TFInteractionArrayI kit 115

MA5011 TF-TFInteractionArrayII kit 115

MA5012 TF-TFInteractionArrayIII kit 115

MA5041 PPARa-TFInteractionArrayI kit 118

MA5042 PPARa-TFInteractionArrayII kit 118

MA5043 PPARa-TFInteractionArrayIII kit 118

MA5044 PPARg-TFInteractionArrayI kit 118

MA5045 PPARg-TFInteractionArrayII kit 118

MA5046 PPARg-TFInteractionArrayIII kit 118

MA5051 NFkBp50-TFInteractionArrayI kit 118

MA5052 NFkBp50-TFInteractionArrayII kit 118

MA5053 NFkBp50-TFInteractionArrayIII kit 118

MA5054 NFkBp65-TFInteractionArrayI kit 118

MA5055 NFkBp65-TFInteractionArrayII kit 118

MA5056 NFkBp65-TFInteractionArrayIII kit 118

MA5110 TF-TFInteractionIRefill kit 115

MA5111 TF-TFInteractionIIRefill kit 115

MA5112 TF-TFInteractionIIIRefill kit 115

MA6110 HumanCytokineAntibodyArray1.0 2membranes 134






MA6310 HumanAngiogenesisAntibodyArray kit 133

MA6320 MouseAngiogenesisAntibodyArray kit 133

MA6410 MouseCytokineAntibodyArray1.0 4membranes 134

MA6412 MouseCytokineAntibodyArray1.0 8membranes 134

ME0001 PPARaCMVExpressionVector 15µg 114

ME0002 PPARb(PPARΔ)CMVExpressionVector 15µg 114

ME0003 PPARg-1CMVExpressionVector 15µg 114

ME0004 PPARg-2CMVExpressionVector 15µg 114

ME0005 NFkBp50CMVExpressionVector 15µg 114

ME0006 NFkBp65(RelA)CMVExpressionVector 15µg 114

ME0007 GR(NR3C1)CMVExpressionVector 15µg 114

ME0010 E2F2CMVExpressionVector 15µg 114

ME0014 IRF1CMVExpressionVector 15µg 114

ME0018 MAFKCMVExpressionVector 15µg 114

ME0019 MAXCMVExpressionVector 15µg 114

ME0020 CITED1CMVExpressionVector 15µg 114

ME0022 NF-YCCMVExpressionVector 15µg 114




ME0024 NR1I2CMVExpressionVector 15µg 114

ME0025 NR1I3CMVExpressionVector 15µg 114

ME0029 PREBCMVExpressionVector 15µg 114

ME0032 SRYCMVExpressionVector 15µg 114

ME0033 STAT1CMVExpressionVector 15µg 114

ME0035 STAT5BCMVExpressionVector 15µg 114

ME0040 USF1CMVExpressionVector 15µg 114

ME0043 ERaCMVExpressionVector 15µg 114

ME0044 FOSCMVExpressionVector 15µg 114

ME0045 JUNCMVExpressionVector 15µg 114

ME0046 NF-ATC1CMVExpressionVector 15µg 114

ME0047 p53CMVExpressionVector 15µg 114

ME0049 GATA1CMVExpressionVector 15µg 114

ME0052 MYCCMVExpressionVector 15µg 114

ME0054 PBX1CMVExpressionVector 15µg 114

ME0055 ATF3CMVExpressionVector 15µg 114

ME0059 ATF1CMVExpressionVector 15µg 114

ME0060 CDX2CMVExpressionVector 15µg 114

ME0061 NR0B1CMVExpressionVector 15µg 114

ME0062 HNF4gCMVExpressionVector 15µg 114

ME0065 BTG2CMVExpressionVector 15µg 114

ME0066 GFI1CMVExpressionVector 15µg 114

ME0071 SMAD1(MADH1)CMVExpressionVector 15µg 114

MP1100 PromoterMethylationPCRKit kit 24

MP1101 14-3-3 ea 24

MP1102 14-3-3 ea 24

MP1103 14-3-3 ea 24

MP1104 14-3-3 ea 24

MP1105 ABL1 ea 24

MP1106 ABL1 ea 24

MP1107 BAGE ea 24

MP1108 BAGE ea 24

MP1109 BAGE ea 24

MP1110 BAGE ea 24

MP1111 BAGE ea 24

MP1112 BAGE ea 24

MP1113 BRCA1 ea 24

MP1114 BRCA1 ea 24

MP1115 BRCA1 ea 24

MP1116 BRCA1 ea 24

MP1117 CalcitoninCGPR ea 24




MP1121 CD14 ea 24

MP1122 CD14 ea 24

MP1123 CDKN2A ea 24

MP1124 CDKN2A ea 24

MP1125 CDKN2A ea 24

MP1126 CDKN2A ea 24

MP1127 COX2 ea 24

MP1128 COX2 ea 24

MP1129 CyclinD2 ea 24




MP1133 DAPK ea 24

MP1134 DAPK ea 24

MP1135 DAPK ea 24

MP1136 DAPK ea 24

MP1137 glut4 ea 24

MP1138 glut4 ea 24

MP1139 H-ras ea 24

MP1140 H-ras ea 24

MP1141 H-ras ea 24

MP1142 H-ras ea 24

MP1143 INFg ea 24

MP1144 INFg ea 24

MP1145 GAGE1 ea 24

MP1146 GAGE1 ea 24

MP1147 GAGE1 ea 24

MP1148 GAGE1 ea 24

MP1149 HOXA2 ea 25

MP1150 HOXA2 ea 25

MP1151 IL4 ea 25

MP1152 IL4 ea 25

MP1153 IRF7 ea 25

MP1154 IRF7 ea 25




MP1155 NIS ea 25

MP1156 NIS ea 25

MP1157 NIS ea 25

MP1158 NIS ea 25

MP1159 NME2 ea 25

MP1160 NME2 ea 25

MP1161 NME2 ea 25

MP1162 NME2 ea 25

MP1163 PDGFB ea 25

MP1164 PDGFB ea 25

MP1165 POMC ea 25

MP1166 POMC ea 25

MP1167 NES ea 25

MP1168 NES ea 25

MP1169 NES ea 25

MP1170 NES ea 25

MP1171 PRA2 ea 25

MP1172 PRA2 ea 25

MP1173 TFF1 ea 25

MP1174 TFF1 ea 25

MP1175 VHL ea 25

MP1176 VHL ea 25

MP1177 Survivin ea 25

MP1178 Survivin ea 25

MP1179 CASP8 ea 25

MP1180 CASP8 ea 25

MP11XX PCRPrimers(forPromoterMethylationPCRKit) ea 24

RC0002 NFkBA549ReporterStableCellLine vial 122

RC0004 STAT-1HeLaReporterStableCellLine vial 122

RC0005 NFATHeLaReporterStableCellLine vial 122

RC0006 AP-1293ReporterStableCellLine vial 122

RC0007 CREB293ReporterStableCellLine vial 122

RC0008 CREBCHOReporterStableCellLine vial 122

RC0009 NFATK562ReporterStableCellLine vial 122

RC0012 SRFHeLaReporterStableCellLine vial 122

RC0013 NFkBHeLaReporterStableCellLine vial 122

RC0014 NFkB293ReporterStableCellLine vial 122

RC0015 NFkBNIH3T3ReporterStableCellLine vial 122

RC0016 NFkBC2C12ReporterStableCellLine vial 122

RC0017 HIFNIH3T3ReporterStableCellLine vial 122

RC0018 GR293ReporterStableCellLine vial 122

RC0019 GRHeLaReporterStableCellLine vial 122

RC1001 TAD-KinaseReporterStableCellLine vial 122

RC2001 CHO/GFP-NFkBp65StableCellLine vial 122

RC2002 CHO/GFP-GRaStableCellLine vial 122

RC5001 NFkBp50KnockdownHeLaStableCellLine vial 122

Product Name Index | Alphabetical


Product Page Product Page

A

14-3-3 24

14-3-3-sigma(SFN)GenePromoterReporterVector 111

ABL1 24

ABL1GenePromoterReporterVector 111

N-Acetyl-L-Cysteine 188

S-AcetylCoenzymeA,LithiumSalt,Ultrapure 188

AcetylatedBSA—SeeAlbumin,Bovine,Acetylated 198

Acrylamide 188

Acrylamide,Ultrapure 188

AcrylamideGelMixesandSolutions

GlycerolTolerantGel(GTG)Buffer,20XSolution 219

RapidGel™40%LiquidAcrylamideStockSolution 237

RapidGel-XL6%DNASequencingGelSolution 238

StopSolution 242

Adenine 188

AdenineSulfate,Dihydrate 188

Adenosine 189

Adenosine-5’-Diphosphate,DisodiumSalt,Dihydrate 189

Adenosine-5’-Monophosphate,DisodiumSalt 189

Adenosine-5’-Monophosphate,FreeAcid 189

Adenosine-5’-Monophosphate,FreeAcid,Monohydrate 189

Adenosine-5’-Triphosphate,100mMSolution,Ultrapure 61,189

Adenosine-5’-Triphosphate,DisodiumSalt 189

S-Adenosyl-L-MethionineSulfatep-Toluenesulfonate 190

Adonitol 190

ADP—SeeAdenosine-5’-Diphosphate,DisodiumSalt,Dihydrate 189

AEBSFHydrochloride 190

AFPELISAKit 134

Agar 190

Agar,Bacteriological,Ultrapure 190

Agar,NobleAgar,Ultrapure 190

Agarose-HE,Ultrapure 192

Agarose-Hi-Res,Separation≤1000bp,Ultrapure 195

Agarose-LE,Ultrapure 194

Agarose-LowMeltSeparation≥1000bp,GeneticPerformanceCertified,Ultrapure 196

Agarose-LowMeltSeparation≤1000bp,GeneticPerformanceCertified,Ultrapure 197

Agarose-ME,Ultrapure 192

Agarose-Separation≥500bp,GeneticPerformanceCertified,Ultrapure 193

Agarose-UltraLowMelt,Ultrapure 197

b-Alanine 198

Albumin,Bovine,Acetylated,Ultrapure 198

Albumin,Bovine,CohnFractionV,pH5.2 198

Albumin,Bovine,Crystallized,pH5.2 198

Albumin,Bovine,pH7 198

Albumin,Bovine,Protease-Free 198

Albumin,Bovine,RIAGrade,pH7 198

Albumin,Egg 199

AlcoholDehydrogenase 199

AlkalinePhosphatases

CalfIntestinalAlkalinePhosphatase(CIAP) 82

ShrimpAlkalinePhosphatase(SAP) 83

SuperSAPShrimpAlkalinePhosphatase 84

4-AminoAntipyrene 199

p-AminoBenzoicAcid(PABA) 199

6-AminoPurine—SeeAdenine 188

L-a-Amino-N-ButyricAcid 199

AminoaceticAcid—SeeGlycine 219

2-AminoethaneSulfonicAcid—SeeTaurine 243

L-a-AminoglutaricAcid—SeeL-GlutamicAcid 218

AmmoniumAcetate,Ultrapure 199

AmmoniumAcetate(NH4OAc),5MSolution,Ultrapure 199

AmmoniumPersulfate(APS),Ultrapure 200

AmmoniumSulfate,Ultrapure 200

AMP—SeeAdenosine-5’-Monophosphate,DisodiumSalt 189

cAMP/CalciumProtein/DNAArray 115

cAMP/CalciumProtein/DNAArrayRefill 115

Ampicillin,SodiumSalt,Ultrapure 200

AMVReverseTranscriptase 90,162

AntibioticG-418Sulfate—SeeG-418Sulfate 217

AP-1293ReporterStableCellLine 122

AP-1ColdProbe 110

AP-1ELISAKit 110

AP-1ELISAKitwithNuclearExtractionKit 110

AP1(1)LuciferaseReporterVector 112


AP3LuciferaseReporterVector 112

APCGenePromoterReporterVector 111

Aprotinin,Ultrapure 200




ARLuciferaseReporterVector 112

L-Arabinose 200

L-(+)-Arabinose 200

L-Arginine 200

L-Arginine,Hydrochloride 201

L-AscorbicAcid 201

L-Asparagine,Anhydrous 201

L-Asparagine,Monohydrate 201

L-AsparticAcid 201

L-AsparticAcid,MagnesiumSalt,Dihydrate 201

L-AsparticAcid,SodiumSalt,Monohydrate 201

ATF1CMVExpressionVector 114

ATF2GenePromoterReporterVector 111


ATP—SeeAdenosine-5’-Triphosphate 189

Avidin-AlkalinePhosphataseConjugate—SeeStreptavidin-AlkalinePhosphataseConjugate 242

B

Bacitracin 202

BacterialCultureMedia

LBAgar,Ready-MadePowder 224

LBBroth,Ready-MadePowder 224

LuriaAgar,Ready-MadePowder 225

LuriaBroth,Ready-MadePowder 225

NZCYMBroth,Ready-MadePowder 231

TerrificBroth,Ready-MadePowder 245

2XYTBroth 251

BacteriologicalPeptone,Ultrapure 202

BAGE 24

BCIP—See5-Bromo-4-Chloro-3-IndolylPhosphate 203

Betaine,5MSolution,Ultrapure 202

Bicine 202

BindingProteins

RecAProtein 65

Single-StrandedDNABindingProtein(SSB) 65

T4Gene32Protein 66

Biotin-11-dXTPAnalog(DNALabelingReagent,DLR) 57

Biotin-11-ψTPAnalog(RNALabelingReagent,RLR) 57

Bis-Acrylamide—SeeN,N’-Methylene-bis-Acrylamide 228

Bis-Tris,Ultrapure 202

BlottingReagents

Denhardt’sSolution,50X 209

SSC,20XSolution 241

SSPE,20XSolution 241

TBS,20XSolution,pH7.4 244

TBSwithTween(TBST),20XSolution,pH7.4 244

BoricAcid,Ultrapure 202

BRCA1 24

BRCA1GenePromoterReporterVector 111

BrewersYeast—SeeYeast,Brewers 251

BrilliantBlueG,Ultrapure 203

BrilliantBlueR,Ultrapure 203

5-Bromo-4-Chloro-3-5-Bromo-4-Chloro-3-IndolylPhosphate,DisodiumSalt,Ultrapure 203

5-Bromo-4-Chloro-3-Indolyl-a-D-Galactopyranoside,Ultrapure 203

5-Bromo-4-Chloro-3-Indolyl-b-D-Galactopyranoside,Ultrapure 203

5-Bromo-4-Chloro-3-IndolylBromocresolGreen,SodiumSalt,Ultrapure 203

BromophenolBlue,SodiumSalt,Ultrapure 204

BSA—SeeAlbumin,Bovine 198

BTG2CMVExpressionVector 114

Buffers


HEPES,1MSolution,pH7.3 222

PBS,10XSolution,pH7.4 232

RapidRunAgaroseBuffer,20XSolution 238

TAEBuffer,10XSolution 243


TBEBuffer,5XSolution 243

TBEBuffer,10XReady-MixedPowder 243

TEBuffer,1XSolution 244

TEBuffer,1XSolution,pH7.0 244

TEBuffer,1XSolution,pH7.0,LowEDTA 244



TEBuffer,50XSolution 244

Tris-Glycine(TG)Buffer,10XSolution 247

Tris-Glycine-SDS(TGS)Buffer,10XSolution 247

Tris/HCl,1MSolution,pH7.0 248






C

CA15-3ELISAKit 134

CA19-9ELISAKit 134

CA125ELISAKit 134

CacodylicAcid,SodiumSaltTrihydrate 204

Caffeine,Anhydrous,Ultrapure 204

CalciumChloride,Dihydrate 204

D-CalciumPantothenate 204

CalcitoninCGPR 24


CancerCellIsolationKit(2) 101



Carbenicillin,DisodiumSalt 205

CarboxypeptidaseY 205

Casein,Hammarsten,Ultrapure 205

Casein(HighNitrogen) 205

CaseinHydrolysate,EnzymaticDigest,Ultrapure 205

Casein,Sodium 205

Casein-Vitafree,Ultrapure 205

CASP8 25

Catalase 205

CD14 24

CDKN2A 24

CDX2CMVExpressionVector 114

CellGrowthProtein/DNAArray 115

CellGrowthProtein/DNAArrayRefill 115

CellularManipulation

DeliverXPeptideTransfectionKits 99

CancerCellIsolationKit 101

Cellulase 205

CesiumChloride,Ultrapure 206

CetylTrimethylAmmoniumBromide 206

CFTRGenePromoterReporterVector 111

CGPR(CALCR)GenePromoterReporterVector 111

Change-ITMultipleMutationSiteDirectedMutagenesisKit 126

ChickenIntestineAlkalinePhosphastase 206

Chloramphenicol,Ultrapure 206

CHO/GFP-GRaStableCellLine 122

CHO/GFP-NFkBp65StableCellLine 122

Cholesterol 206

CholineChloride 207

CIITAGenePromoterReporterVector 111

CITED1CMVExpressionVector 114

CitricAcid,Anhydrous 207

CitricAcid,TrisodiumSalt,Dihydrate,Ultrapure 207

Cloning

Ligate-ITKit 125

Cocarboxylase 207

CoenzymeI—Seeb-NicotinamideAdenineDinucleotide 229

CoenzymeII—SeeNicotinamideAdenineDinucleotidePhosphate 230

CoenzymeA,FreeAcid,Trihydrate 207

CoenzymeA,TrilithiumSalt,Dihydrate 207

Collagen 207

Collagenase,TypeI 207

ComboProtein/DNAArray 116

ComboProtein/DNARefillKit 116

CompactReactionColumns 207-208

ControlHEK292PMANuclearExtract 110

CoomassieStains—SeeBrilliantBlueG&R 203

COX2 24

COX2(PTGS2)GenePromoterReporterVector 111

CRCLowerFilters 208

CRCUpperFilters 208

CRE(1)LuciferaseReporterVector 112

CreatineKinase 208

CreatinePhosphate,DisodiumSalt,Tetrahydrate 208

CREB293ReporterStableCellLine 122

CREBCHOReporterStableCellLine 122

CTAB—SeeCetylTrimethylAmmoniumBromide 206

CTP—SeeCytidine-5’-Triphosphate 209

Cyanocobalamin—SeeVitaminB12 250

CyclinD2 24

a-Cyclodextrin 208

L-Cystine 208

L-Cysteine,Hydrochloride,Monohydrate 208

Cytidine,FreeBase 208

Cytidine-5’-Triphosphate,DisodiumSalt,Dihydrate 209

CytochromeC 209




D

DAPI 209

DAPK 24

7-deaza-2’-Deoxyguanosine-5’-Triphosphate,LithiumSalt,10mMSolution 60

DeliverXsiRNABuffer-1 98

DeliverXsiRNABuffer-2 98

DeliverXsiRNATransfectionEvaluationKit 98

DeliverXsiRNATransfectionKit 98

DeliverXPeptideBuffer-1 99

DeliverXPeptideBuffer-2 99

DeliverXPeptideTransfectionEvaluationKit 99

DeliverXPeptideTransfectionKit 99

DeliverXPlussiRNABuffer-1 98

DeliverXPlussiRNABuffer-2 98

DeliverXPlussiRNATransfectionEvaluationKit 98

DeliverXPlussiRNATransfectionKit 98

DeliverXPre-formedPeptideTransfectionComplexControl 99

DeliverXTAMRA-labeledPeptideControl 99

Denhardt’sSolution,50X,Ultrapure 209

2’-Deoxyadenosine 209

2’-Deoxyadenosine-5’-Triphosphate,SodiumSalt,100mMSolution,Ultrapure 59,210

DeoxycholicAcid,SodiumSalt 210

2’-Deoxycytidine,Hydrochloride,Synthetic 210

2’-Deoxycytidine-5’-Triphosphate,SodiumSalt,100mMSolution,Ultrapure 59,210

2’-Deoxyguanosine 210

2’-Deoxyguanosine,MonohydrateSynthetic 210

7-deaza-2’-Deoxyguanosine-5’-Triphosphate,LithiumSalt,10mMSolution 210

2’-Deoxyguanosine-5’-Triphosphate,SodiumSalt,100mMSolution,Ultrapure 59,210

2’-Deoxynucleoside-5’-TriphosphateMixes—SeePCRNucleotideMixes 232

2’-Deoxynucleoside-5’-Triphosphates,SodiumSalt,100mMSolutions, 59,211SetsofFour,Ultrapure,4x25µmol(250µl)eachdNTPperpack

2’-Deoxynucleoside-5’-Triphosphates,SodiumSalt,100mMSolutions, 59,211SetsofFour,Ultrapure,4x100µmol(1ml)eachdNTPperpack

2’-Deoxynucleoside-5’-Triphosphates,SodiumSalt,100mMSolutions, 59,211SetsofFour,Ultrapure,4x500µmol(5ml)eachdNTPperpack

2-Deoxy-D-Ribose,Ultrapure 211

2’-Deoxythymidine-5’-Triphosphate,SodiumSalt,100mMSolution,Ultrapure 59,211

2’-Deoxyuridine-5’-Triphosphate,SodiumSalt,100mMSolution,Ultrapure 59,211

DEPC—SeeDiethylPyrocarbonate 212

Dextran,Ultrapure 211

DextranSulfate,SodiumSalt 211

DextranSulfate,SodiumSalt,Ultrapure 211

D-(+)-Dextrose,Anhydrous 211

D-(+)-Dextrose,Monohydrate 212

Diaphorase 212

2’,3’-Dideoxyadenosine-5’-Triphosphate,Ultrapure,10mMaqueoussolution, 60,212pH7.5

2’,3’-Dideoxycytidine-5’-Triphosphate,Ultrapure,10mMaqueoussolution, 60,212pH7.5

2’,3’-Dideoxynucleoside-5’-Triphosphates,10mMSolutions,SetsofFour, 60,212Ultrapure,4x0.5µmol(50µl)eachddNTPperpack

2’,3’-Dideoxynucleoside-5’-Triphosphates,10mMSolutions,SetsofFour, 60,212Ultrapure,4x1.0µmol(100µl)eachddNTPperpack

2’,3’-Dideoxynucleoside-5’-Triphosphates,100mMSolutions,SetofFour, 60,212Ultrapure,4x4µmol(40µl)eachddNTPperpack

2’,3’-Dideoxythymidine-5’-Triphosphate,Ultrapure,10mMaqueoussolution, 60,212pH7.5

DiethylPyrocarbonate,Ultrapure 212

DihydrostreptomycinSulfate 213

Dimedone 213

p-DimethylaminoBenzaldehyde 213

p-DimethylaminoCinnamaldehyde 213

Dithioerythritol,Ultrapure 213

Dithiothreitol 213

Dithiothreitol,Ultrapure 213

Dithiothreitol(DTT),0.1MSolution,Ultrapure 213

DNA

DNA,CalfThymus 128

DNA,E. coli 128

DNA,FishSperm 128

DNA,FishSperm,Activated 128

DNA,FishSperm,SodiumSalt 128

DNA-Cellulose,Double-Stranded 128

DNA-Cellulose,Single-Stranded 129

M13mp18,Single-Stranded,0.2µg/ul 129

M13mp18DNA,Single-Stranded,1.0µg/µl 129

Oligo(dT)12-18Primer,MWa4500 129

Oligop(dT)12-18SodiumSalt 129

pUC19DNA,0.2µg/µl 129


DNACleanup

PrepEaseDNACleanupKits 146

DNALadder,1kbPlus 131

DNALadder,100bp 130




DNALoadingBuffer(OXG),6X 130

DNALoadingBuffer(BXF),6X 131

DNAMarkers

DNALadder,1kbPlus 131

DNALadder,100bp 130

PCRMarkers,50-2000bp 130

DNAPolymeraseI 87

DNAPolymerases

DNAPolymeraseI 87

Exonuclease-FreeKlenow 86

Exonuclease-FreeKlenowTrisBuffer 86

KlenowDNAPolymeraseI 87

SequenaseVersion2.0DNAPolymerase 88,174

T7DNAPolymerase 88

TthDNAPolymerase 89

DNA-Cellulose,Double-Stranded 124

DNA-Cellulose,Single-Stranded 124

DNaseI 213

DNaseI,Lyophilized 72

DNaseI,Solution 72

rDNaseI,RNase-Free 73

10XDNaseIBuffer 74

DNases

10XDNaseIBuffer 74


DNaseI,Solution 72


ShrimpDNase,Recombinant 74

DodecylSulfate,LithiumSalt—SeeLithiumDodecylSulfate 225

DodecylSulfate,SodiumSalt—SeeSodiumDodecylSulfate 240

N-DodecylTrimethylammoniumBromide 214

DPN—SeeNicotinamideAdenineDinucleotide 229

DTAB—SeeN-DodecylTrimethylammoniumBromide 214

DTE—SeeDithioerythritol 213

DTT—SeeDithiothreitol 213

E

E2F-1ELISAKit 110

E2F-1ELISAKitwithNuclearExtractionKit 110

E2F(1)LuciferaseReporterVector 112

E2F2CMVExpressionVector 114


E2F1(3)LuciferaseReporterVector 112

E. coliDNALigase 70

E. coliRNAPolymeraseHoloenzyme 91,159

EDTA—SeeEthylenediamineTetraaceticAcid 215

EggWhite—SeeAlbumin-Egg 199

EGTA—SeeEthyleneGlycol-o-o’-bis(2-aminoethyl)-N,N,N’,N’,-TetraaceticAcid 215

Elastase 214

ELK1LuciferaseReporterVector 112

EMSAComboKit(anythreeprobes) 103

EMSAKit-Seepage104forfulllisting 103

EMSAKitwithoutProbeSet 103

EMSATFProbeSets-Seepage104forfulllisting 103

EndPointPCR

RubyTaqDNAPolymerase 13

RubyTaqPCRMasterMix(2X) 14

TaqDNAPolymerase 11

TaqPCRKit 12

TaqPCRMasterMix(2X) 12

Endonucleases

HumanApurinic/ApyrimidinicEndonuclease1(APE1) 75

T4EndonucleaseVII 75

Enolase 214

ERControlMCF7NuclearExtract 110

ERLuciferaseReporterVector 112

ERaCMVExpressionVector 114

ERaELISAKit 110

ERaELISAKitwithNuclearExtractionKit 110

ERaGenePromoterReporterVector 111

EREColdProbe 110

Ergocalciferol 214

Esculin 214

EthidiumBromide,Ultrapure 214

EthidiumBromideDrops,Ultrapure 214

EthylenediaminetetraaceticAcid,(EDTA),0.5MSolution,Ultrapure 215

EthylenediaminetetraaceticAcid,(EDTA)DisodiumSalt,Dihydrate 215

EthylenediaminetetraaceticAcid,(EDTA)DisodiumSalt,Dihydrate,Ultrapure 215

EthyleneDiamineTetraaceticAcid,(EDTA)TetrasodiumSalt,Dihydrate,Ultrapure 215

EthyleneGlycol-O,O’-Bis(2Amino-ethyl)-N,N,N’,N’,-TetraaceticAcid(EGTA),Ultrapure 215




ExonucleaseI,Highconcentration,20units/µl 76

ExonucleaseI,Standardconcentration,10units/µl 76

ExonucleaseIII 77

ExonucleaseVII 77


Exonucleases

ExonucleaseI 76

ExonucleaseIII 77

ExonucleaseVII 77

T7Gene6Exonuclease 78

ExoSAP-ITPCRProductCleanup 49,148

F

FAM-labeledsiRNAControl 98

FetalBovineSerum(FBS) 215

FHITGenePromoterReporterVector 111

FideliTaqDNAPolymerase 15

FideliTaqPCRMasterMix(2X) 16

FideliTaqRT-PCRMasterMix(2X) 46

First-StrandcDNASynthesisKitforReal-TimePCR 39

FKHRELISAKit 110

FKHRELISAKitwithNuclearExtractionKit 110

FlavinAdenineDinucleotide,DisodiumSalt,Hydrate 216

FluoresceinPassiveReferenceDye 53

FolicAcid 216

Formamide,Ultrapure 216

FOSCMVExpressionVector 114

b-D-Fructose 216

D-Fructose-6-Phosphate,DisodiumSalt 216

G

G-418Sulfate,Ultrapure 217

G6PDGenePromoterReporterVector 111

GAGE1 24

GAGE1GenePromoterReporterVector 111

D-(+)-Galactose 217

GAS/ISRELuciferaseReporterVector 112

GASLuciferaseReporterVector 112

GATALuciferaseReporterVector 112

GATA-1ColdProbe 110

GATA-1ControlRecombinantProtein 110

GATA-1ELISAKit 110

GATA-1ELISAKitwithNuclearExtractionKit 110

GATA1CMVExpressionVector 114

GATA1LuciferaseReporterVector 112

GATA1/GATA2LuciferaseReporterVector 112




GDP—SeeGuanosine-5’-Diphosphate 221

GelElectrophoresisReagents

EthidiumBromideDrops 214


RapidRunAgaroseBuffer,20XSolution 238



TBEBuffer,5XSolution 243

TBEBuffer,10XReady-MixedPowder 243

Tris-Glycine(TG)Buffer,10XSolution 247

Tris-Glycine-SDS(TGS)Buffer,10XSolution 247

GelMixesandSolutions


RapidGel40%LiquidAcrylamideStockSolution 237

RapidGel-XL6%DNASequencingGelSolution 238

StopSolution 242

GelExtraction

PrepEaseGelExtractionKits 146

GenePromoterReporterVectors

14-3-3-sigma(SFN)GenePromoterReporterVector 111

ABL1GenePromoterReporterVector 111

APCGenePromoterReporterVector 111

ATF2GenePromoterReporterVector 111

BRCA1GenePromoterReporterVector 111

CFTRGenePromoterReporterVector 111

CGPR(CALCR)GenePromoterReporterVector 111

CIITAGenePromoterReporterVector 111

COX2(PTGS2)GenePromoterReporterVector 111

ERaGenePromoterReporterVector 111

FHITGenePromoterReporterVector 111

G6PDGenePromoterReporterVector 111

GAGE1GenePromoterReporterVector 111

hMLH1GenePromoterReporterVector 111




HOXA2GenePromoterReporterVector 111


IFNgGenePromoterReporterVector 111

IGRPGenePromoterReporterVector 111

IL1aGenePromoterReporterVector 111

IL2GenePromoterReporterVector 111


IRF7GenePromoterReporterVector 111

JUNBGenePromoterReporterVector 111

MDR1GenePromoterReporterVector 111

MyoDGenePromoterReporterVector 111

POMCGenePromoterReporterVector 111

RB1GenePromoterReporterVector 111

STAT1GenePromoterReporterVector 111

SURVIVINGenePromoterReporterVector 111

THBS2GenePromoterReporterVector 111

TIMP3GenePromoterReporterVector 111

TNFaGenePromoterReporterVector 111

VHLGenePromoterReporterVector 111

v-mycGenePromoterReporterVector 111

GeneticinDisulfate—SeeG-418Sulfate 217

GenomicDNAIsolation

PrepEaseGenomicDNAIsolationKit 147

GentamicinSulfate 217

GFI1CMVExpressionVector 114

g-Globulins,FractionII 217

a-D-(+)-Glucose—SeeDextrose 211

GlucoseOxidase 217

D-Glucose-6-Phosphate,DisodiumSalt,Dihydrate 217

Glucose-6-PhosphateDehydrogenase 217,218

Glucose-6-PhosphateGlutamateDehydrogenase 218

glut4 24

L-GlutamicAcid 218

L-GlutamicAcid,MonosodiumSalt,Monohydrate 218

L-Glutamine 218

GlutamylCysteinylGlycine—SeeGlutathione 218

Glutathione-Oxidized 218

Glutathione-Reduced 218

Glycerin—SeeGlycerol 219

Glycerol,Ultrapure 219

GlycerolTolerantGel(GTG)Buffer,20XSolution,Ultrapure 219

b-Glycerophosphate,SodiumSalt,Pentahydrate 219

Glycine 220

Glycine,Ultrapure 219

Glycogen,20mg/ml,Ultrapure 220

Glycogen,Ultrapure 220

Glycosylases

Uracil-DNAGlycosylase,E. coli 67

Uracil-DNAGlycosylase,Heat-Labile 67

Glycyl-Glycine 220

GR293ReporterStableCellLine 122

GRColdControlProbe 110

GRControlHela(DEX)NuclearExtract 110

GRELISAKit 110

GRELISAKitwithNuclearExtractionKit 110

GRHeLaReporterStableCellLine 122

GRLuciferaseReporterVector 112

GR(NR3C1)CMVExpressionVector 114

GST-TaggedProteinPurification

PrepEaseProteinPurificationGlutathioneAgarose4B 155

GTP—SeeGuanosine-5’-Triphosphate 221

GuanidineHydrochloride 221

GuanidineHydrochloride,Ultrapure 220

GuanidineThiocyanate,Ultrapure 221

Guanosine-5’-Diphosphate,DisodiumSalt 221

Guanosine-5’-Triphosphate,DisodiumSalt 221

Guanosine-5’-Triphosphate,SodiumSalt,100mMSolution,Ultrapure 61,221

H

Hemin 221

Heparin,SodiumSalt 221

HEPES,1MSolution,pH7.3,Ultrapure 222

HEPES,SodiumSalt 222

HEPES,Ultrapure 222

Hexokinase 222

HIFColdControlProbe 110

HIFControlCos1NuclearExtract 110

HIFNIH3T3ReporterStableCellLine 122

HIF-1aELISAKit 110

HIF-1aELISAKitwithNuclearExtractionKit 110




HIF1LuciferaseReporterVector 112

Histidine-TaggedProteinPurificationKits

PrepEaseHistidine-TaggedMaxiKit–HighSpecificity 153

PrepEaseHistidine-TaggedMaxiKit–HighYield 154

PrepEaseHistidine-TaggedMidiKit–HighSpecificity 153

PrepEaseHistidine-TaggedMidiKit–HighYield 154

PrepEaseHistidine-TaggedMiniKit–HighSpecificity 153

PrepEaseHistidine-TaggedMiniKit–HighYield 154

Histidine-TaggedProteinPurificationResins

PrepEaseHistidine-TaggedHighSpecificityPurificationResin 155

PrepEaseHistidine-TaggedHighYieldPurificationResin 155

L-Histidine,FreeBase 222

L-Histidine,Monohydrochloride,Monohydrate 222

hMLH1GenePromoterReporterVector 111

HNF-3bELISAKit 110

HNF-3bELISAKitwithNuclearExtractionKit 110

HNF4gCMVExpressionVector 114

HotStartPCR

HotStart-ITBindingProtein 21

HotStart-ITTaqDNAPolymerase 19

HotStart-ITTaqPCRMasterMix(2X) 20

HotStart-ITBindingProtein 21

HotStart-ITFideliTaqDNAPolymerase 17,25

HotStart-ITFideliTaqPCRMasterMix(2X) 18

HotStart-ITProbeOne-StepqRT-PCRMasterMixKit 44

HotStart-ITProbeqPCRMasterMix(2X) 38

HotStart-ITProbeqPCRMasterMixwithUDG(2X) 38

HotStart-ITSYBRGreenOne-StepqRT-PCRMasterMixKit 44

HotStart-ITSYBRGreenqPCRMasterMix(2X) 32

HotStart-ITSYBRGreenqPCRMasterMixwithUDG(2X) 32

HotStart-ITTaqDNAPolymerase 19

HotStart-ITTaqPCRMasterMix(2X) 20

HOXA2 25



H-ras 24

HSELuciferaseReporterVector 112

HTExoSAP-ITHigh-ThroughputPCRProductCleanup 50,149

HumanAngiogenesisAntibodyArray 133


HumanCytokineAntibodyArray1.0 134

HumanCytokineAntibodyArray3.0 134

HumanGAPDHsiRNAControl 98

L-HydroxysuccinicAcid—SeeL-MalicAcid 227

I

IFNgGenePromoterReporterVector 111

IGRPGenePromoterReporterVector 111

IL1aGenePromoterReporterVector 111


IL4 25


Imidazole,Ultrapure 223

INFg 24

IPTG—SeeIsopropyl-b-D-Thiogalactopyranoside 223

IRF1CMVExpressionVector 114

IRF1LuciferaseReporterVector 112

IRF7 25

IRF7GenePromoterReporterVector 111

IsocitrateDehydrogenase,(NADP+Specific),Recombinant 223

DL-Isoleucine 223

L-Isoleucine 223

Isopropyl-b-D-Thiogalactopyranoside(IPTG),DioxaneFree,Ultrapure 223

ISRELuciferaseReporterVector 112

J

JUNCMVExpressionVector 114

JUNBGenePromoterReporterVector 111

K

KanamycinSulfate 223

a-KetoglutaricAcid,DisodiumSalt,Dihydrate 224

a-KetoglutaricAcid,FreeAcid 224

Kinases

OptiKinase 68

T4PolynucleotideKinase(PNK) 69


L

Labeling

SequenaseRandomPrimerLabelingKit 127

L-(+)-LacticAcid,LithiumSalt 224

Lactoferrin 224




D-(+)-Lactose,Monohydrate 224

N-LauroylSarcosine,SodiumSalt,Ultrapure 224

LaurylSulfate,LithiumSalt—SeeLithiumDodecylSulfate 225

LaurylSulfate,SodiumSalt—SeeSodiumDodecylSulfate 240

LBAgar,Ready-MadePowder 224

LBBroth,Ready-MadePowder 224

Lecithin 224

LeptomycinB 224

DL-Leucine 225

L-Leucine 225

Leupeptin,Ultrapure 225

Ligases

E. coliDNALigase 70

T4DNALigase 70

T4RNALigase 71

T4RNALigase2 71

Ligate-ITRapidLigationKit 125

DL-a-LipoicAcid 225

LithiumChloride(LiCl),7.5MSolution,Ultrapure 225

LithiumDodecylSulfate,Ultrapure 225

LithiumLactate—SeeL-(+)-LacticAcid,LithiumSalt 224

LithiumLaurylSulfate—SeeLithiumDodecylSulfate 225

LongandAccuratePCR

FideliTaqDNAPolymerase 15

FideliTaqPCRMasterMix(2X) 16

HotStart-ITFideliTaqDNAPolymerase 17

HotStart-ITFideliTaqPCRMasterMix(2X) 18

LowMolecularWeightMarker,10-100nt 132,165

LuciferaseReporterVectors

TA-luc(EmptyControl)LuciferaseReporterVector 112



AP3LuciferaseReporterVector 112

ARLuciferaseReporterVector 112

CRE(1)LuciferaseReporterVector 112



E2F1(3)LuciferaseReporterVector 112

ELK1LuciferaseReporterVector 112

ERLuciferaseReporterVector 112

GAS/ISRELuciferaseReporterVector 112

GASLuciferaseReporterVector 112

GATALuciferaseReporterVector 112

GATA1/GATA2LuciferaseReporterVector 112





GRLuciferaseReporterVector 112

HIF1LuciferaseReporterVector 112

HSELuciferaseReporterVector 112

IRF1LuciferaseReporterVector 112

ISRELuciferaseReporterVector 112

MEF1LuciferaseReporterVector 112



NF-ATLuciferaseReporterVector 112

NFkB(1)LuciferaseReporterVector 112


p53LuciferaseReporterVector 112

PRLuciferaseReporterVector 112

RARLuciferaseReporterVector 112

RXRLuciferaseReporterVector 112

RXR(2)LuciferaseReporterVector 112

SmadLuciferaseReporterVector 112

Smad3/Smad4LuciferaseReporterVector 112

Sp1LuciferaseReporterVector 112

SP1(2)LuciferaseReporterVector 112

SRELuciferaseReporterVector 112

SRFLuciferaseReporterVector 112

STAT1p84/p91LuciferaseReporterVector 112

STAT1(2)LuciferaseReporterVector 112


STAT4LuciferaseReporterVector 112

VDRLuciferaseReporterVector 112

YY1LuciferaseReporterVector 112

LuriaAgar,Ready-MadePowder 225

LuriaBroth,Ready-MadePowder 225

L-Lysine,Hydrochloride 226

Lysozyme,Ultrapure 226




M

M13mp18,Single-Stranded,0.2µg/ul 129

M13mp18DNA,Single-Stranded,1.0µg/µl 129

MAFKCMVExpressionVector 114

MagnesiumChloride(MgCl2),1MSolution,Ultrapure 226

MagnesiumChloride,Hexahydrate,Ultrapure 226

MagniTaqMultiplexPCRMasterMix(2X) 22

MalateDehydrogenase,Lyophilized 226

MaleicAcid 226

DL-MalicAcid 226

L-MalicAcid 227

Maltose,Monohydrate 227

D-(+)-Maltose,Monohydrate 227

MammalianFull-lengthExpressionVectors



BTG2CMVExpressionVector 114

CDX2CMVExpressionVector 114

CITED1CMVExpressionVector 114

E2F2CMVExpressionVector 114

ERaCMVExpressionVector 114

FOSCMVExpressionVector 114

GATA1CMVExpressionVector 114

GFI1CMVExpressionVector 114

GR(NR3C1)CMVExpressionVector 114

HNF4gCMVExpressionVector 114

IRF1CMVExpressionVector 114

JUNCMVExpressionVector 114

MAFKCMVExpressionVector 114

MAXCMVExpressionVector 114

MYCCMVExpressionVector 114

NF-ATC1CMVExpressionVector 114

NFkBp50CMVExpressionVector 114

NFkBp65(RelA)CMVExpressionVector 114

NF-YCCMVExpressionVector 114

NR0B1CMVExpressionVector 114

NR1I2CMVExpressionVector 114


p53CMVExpressionVector 114

PBX1CMVExpressionVector 114

PPARaCMVExpressionVector 114

PPARb(PPARΔ)CMVExpressionVector 114

PPARg-1CMVExpressionVector 114


PREBCMVExpressionVector 114

SMAD1(MADH1)CMVExpressionVector 114

SRYCMVExpressionVector 114

STAT1CMVExpressionVector 114

STAT5BCMVExpressionVector 114

USF1CMVExpressionVector 114

Mannitol 227

D-(+)-Mannose 227

Markers

DNAMarkers 130

OligonucleotideMarker 132

ProteinMarkers 136

MAXCMVExpressionVector 114

MDR1GenePromoterReporterVector 111


MEF-2ColdControlProbe 110

MEF-2ControlNuclearExtract 110

MEF-2ELISAKit 110

MEF-2ELISAKitwithNuclearExtractionKit 110



MES,Anhydrous 227

MES,Monohydrate,Ultrapure 227

MES,SodiumSalt 227

5-Methyl-2’-Deoxycytidine-5’-Monophosphate,DisodiumSalt 228

MethyleneBlue,Trihydrate 228

N,N’-Methylene-bis-Acrylamide,Ultrapure 228

4-Methylumbelliferyl-a-L-Iduronide,SodiumSalt 228

MicrococcalNuclease 81

MineralOil,Ultrapure 228

M-MLVReverseTranscriptase 90,162

MonosodiumGlutamate—SeeGlutamicAcid,MonosodiumSalt 218

MOPS,Ultrapure 228

MouseAngiogenesisAntibodyArray 133

MouseCytokineAntibodyArray1.0 134




mRNAAssay

Poly(A)Tail-LengthAssayKit 158

MTT,Ultrapure 228

MultiplexPCR

HotStart-ITFideliTaqDNAPolymerase 25

MagniTaqMultiplexPCRMasterMix(2X) 22

PromoterMethylationPCRKit 24

Mutagenesis

Change-ITMultipleMutationSiteDirectedMutagenesisKit 126

MYCCMVExpressionVector 114

MyoDGenePromoterReporterVector 111

N

b-NAD—SeeNicotinamideAdenineDinucleotide 229

NADH—Seeb-NicotinamideAdenineDinucleotide,Reduced 230

NADP—SeeNicotinamideAdenineDinucleotidePhosphate 230

NADPH—SeeNicotinamideAdenineDinucleotidePhosphate,Reduced 230

a-NaphthylPhosphate,MonosodiumSalt 229

NBT—Seep-NitroBlueTetrazoliumChloride 230

Neocuproine,Hydrochloride 229

NeomycinSulfate,Ultrapure 229

NES 25

NeutralRed 229

NextGenerationSequencing(NGS)LibraryPrep

Prep2SeqDNALibraryPrepKitforIllumina 123,170

Prep2SeqMultiplexOligoAdaptersforIlluminaplatform 124,171

NF-ATLuciferaseReporterVector 112

NF-ATC1CMVExpressionVector 114

NFATHeLaReporterStableCellLine 122

NFATK562ReporterStableCellLine 122

NFkB293ReporterStableCellLine 122

NFkBA549ReporterStableCellLine 122

NFkBC2C12ReporterStableCellLine 122

NFkBc-RelELISAKit 110

NFkBc-RelELISAKitwithNuclearExtractionKit 110

NFkBControlNuclearExtract 110

NFkBColdControlProbe 110

NFkBHeLaReporterStableCellLine 122



NFkBNIH3T3ReporterStableCellLine 122

NFkBp50CMVExpressionVector 114

NFkBp50ELISAKit 110

NFkBp50ELISAKitwithNuclearExtractionKit 110

NFkBp50KnockdownHeLaStableCellLine 122

NFkBp50-TFInteractionArrayI 118

NFkBp50-TFInteractionArrayII 118

NFkBp50-TFInteractionArrayIII 118

NFkBp52ELISAKit 110


NFkBp65ELISAKit 110


NFkBp65(RelA)CMVExpressionVector 114

NFkBp65-TFInteractionArrayI 118

NFkBp65-TFInteractionArrayII 118

NFkBp65-TFInteractionArrayIII 118

NFkBRel-bELISAKit 110

NFkBRel-bELISAKitwithNuclearExtractionKit 110

NF-YCCMVExpressionVector 114

Niacinamide 229

Nicotinamide—SeeNiacinamide 229

b-NicotinamideAdenineDinucleotide 229

b-NicotinamideAdenineDinucleotide,LithiumSalt,Dihydrate 229

b-NicotinamideAdenineDinucleotide,MonosodiumSalt 230

b-NicotinamideAdenineDinucleotidePhosphate,Monosodium,Tetrahydrate 230

b-NicotinamideAdenineDinucleotidePhosphate-Reduced,Tetrasodium,Tetrahydrate230

b-NicotinamideAdenineDinucleotide-Reduced,Disodium,Trihydrate 230

NIS 25

p-NitroBlueTetrazoliumChloride,Ultrapure 230

4-Nitrophenyl-N-Acetyl-b-D-Galactosaminide 230

4-Nitrophenyl-a-L-Fucopyranoside 231

4-Nitrophenyl-a-D-Galactopyranoside 231

4-Nitrophenyl-b-D-Glucopyranoside 231

4-Nitrophenyl-b-D-Glucuronide 231

4-Nitrophenyl-a-D-Maltoside 231

4-Nitrophenyl-b-D-Mannopyranoside 231

4-Nitrophenyl-b-D-Xylopyranoside 231

NME2 25

Non-SpecificNucleases


PhosphodiesteraseI 81




NR0B1CMVExpressionVector 114



NuclearExtractionKit 102

NuclearReceptorProtein/DNAArray 115

NuclearReceptorProtein/DNAArrayRefill 115

Nucleases


DNaseI,Solution 72


10XDNaseIBuffer 74

ExonucleaseI 76

ExonucleaseIII 77

ExonucleaseVII 77




RNaseI 79

RnaseI'A',Powder 79

RNaseA,Lyophilized 79

RNaseA,MBGrade,Solution 80

RNaseH 80




Nucleoside-5’-Triphosphates(ATP,CTP,GTP,UTP),SetofFour,100mM 61,231Solutions,Ultrapure,4x25µmol(250µl)eachNTPperpack

NucleotideLabelingReagents

Biotin-11-dXTPAnalog(DNALabelingReagent,DLR) 57

Biotin-11-ψTPAnalog(RNALabelingReagent,RLR) 57

NucleotideSolutions

Adenosine-5’-Triphosphate(ATP) 189

2’-Deoxyadenosine-5’-Triphosphate(dATP) 210

2’-Deoxycytidine-5’-Triphosphate(dCTP) 210

2’-Deoxyguanosine-5’-Triphosphate(dGTP) 210

2’-Deoxynucleoside-5’-Triphosphates(dNTP),SetofFour 211

2’-Deoxythymidine-5’-Triphosphate(dTTP) 211

2’-Deoxyuridine-5’-Triphosphate(dUTP) 211

2’,3’-Dideoxyadenosine-5’-Triphosphate(ddATP) 212

2’,3’-Dideoxycytidine-5’-Triphosphate(ddCTP) 212

2’,3’-Dideoxythymidine-5’-Triphosphate(ddTTP) 212

Guanosine-5’-Triphosphate(GTP) 221

PCR-NucleotideMixes 232

NZCYMBroth,Ready-MadePowder 232

O

Oligo(dT)12-18Primer,MW≅4500 129,231

Oligop(dT)Cellulose,Ultrapure 125

Oligop(dT)12-18SodiumSalt 129,232

OligonucleotideMarker

LowMolecularWeightMarker,10–100nt 132

One-StepRT-PCRKit 47

OptiKinase 68

Ovalbumin—SeeAlbumin,Egg 199

P

p53CMVExpressionVector 114

p53ColdControlProbe 110

p53ELISAKit 110

p53ELISAKitwithNuclearExtractionKit 112

p53LuciferaseReporterVector 112

PABA—Seep-AminoBenzoicAcid 199

Pancreatin1X 232

ParaformaldehydeSolution,4%inPBS 232

PassiveReferenceDyes

FluoresceinPassiveReferenceDye 53

ROXPassiveReferenceDye 53

PBS,10XSolution,pH7.4,Ultrapure 232

PBX1CMVExpressionVector 114

PCRAmplification

EndPointPCR 11

HotStartPCR 19

LongandAccuratePCR 15

MultiplexPCR 22

Real-TimeqPCR 26

Real-TimeReverseTranscriptionPCR(qRT-PCR) 39

ReverseTranscriptionRT-PCR—One-StepMasterMix 45

ReverseTranscriptionRT-PCR—Kits 47

PCRCleanup






PCRMarkers,50-2000bp 130

PCRNucleotideMix,10mMSolution,Ultrapure 60,232

PCRNucleotideMix,25mMSolution,Ultrapure 60,232

PCRNucleotideMixwithdUTP,10mMSolution,Ultrapure 60,232

PCRPrimers(forPromoterMethylationPCRKit) 24

14-3-3 24

ABL1 24

BAGE 24

BRCA1 24

CalcitoninCGPR 24

CASP8 25

CD14 24

CDKN2A 24

COX2 24

CyclinD2 24

DAPK 24

GAGE1 24

glut4 24

H-ras 24

HOXA2 25

IL4 25

INFg 24

IRF7 25

NES 25

NIS 25

NME2 25

PDGFB 25

POMC 25

PRA2 25

Survivin 25

TFF1 25

VHL 25

PCRProductPre-SequencingKit 52

PCRPurification



PCRProductPre-SequencingKit 52

PrepEaseDNACleanupKits 52,146

PrepEaseGelExtractionKits 52,146

PrepEasePCRPurification96-WellPlateKits(Ultrafiltration) 53,148

SBECleanupReagent 52

PDGFB 25

PEG

PolyethyleneGlycol400 235



PenicillinG,Potassium 232

Pepsin 232

Pepsin1:10,000 233

PepstatinA,Ultrapure 233

Peptones

BacteriologicalPeptone 202

CaseinHydrolysate 205

YeastExtractandHydrolysate 251

Phenol,pH8.0,Equilibrated,Ultrapure 233

Phenol,Ultrapure 233

Phenol:Chloroform:IsoamylAlcohol(25:24:1),Ultrapure 233

PhenolRed,SodiumSalt 234

Phenolphthalein 234

Phenolsulfonphthalein—SeePhenolRed 234

DL-Phenylalanine 234

L-Phenylalanine 234

PhenylmethylSulfonylFluoride,Ultrapure 234

Phosphatases




Phosphocreatine—SeeCreatinePhosphate 208


PIPES 208

PIPES,DisodiumSalt 234

PlasmidPurification-BAC

PrepEaseBACPurificationKit 145

PlasmidPurification-Endotoxin-FreePreps

PrepEaseEndotoxin-FreeMaxiPlasmidKit 142

PlasmidPurification-HighThroughput

PrepEasePlasmid96-wellPlateKit 144

PrepeasePlasmid96-wellPlateCoreKit 144




PlasmidPurification-MediumThroughput

PrepEasePlasmid8-wellStripKit 143

PrepEase8-wellStripStarterKitforVacuum 143

PrepEaseVacuumManifold 143

PrepEasePurification-MiditoGigaPreps

PrepEaseMaxiPlasmidKit 140

PrepEaseMidiPlasmidKit 140

PrepEaseGigaPlasmidKit 140

PlasmidPurification-SpinColumns

PrepEaseMiniSpinPlasmidKit 141

PrepEaseQuickMiniSpinPlasmidKit 141

PlasmidPurification-Yeast

PrepEaseYeastPlasmidIsolationKit 145

PMSF—SeePhenylmethylSulfonylFluoride 234

PNK

OptiKinase 68

T4PolynucleotideKinase 69

Poly(A)Polymerase,Yeast 91,159

Poly(A)Tail-LengthAssayKit 158

PolyanetholesulfonicAcid,SodiumSalt 234

Poly(dI-dC)•Poly(dI-dC)SodiumSalt 235


PolyethyleneGlycol6000,Flakes 235

PolyethyleneGlycol8000,Flakes,Ultrapure 235

PolyethyleneGlycol8000,Powder 235

Polymerases


DNAPolymeraseI 87



Exonuclease-FreeKlenowTrisBuffer 86





T3RNAPolymerase 92,160

T7DNAPolymerase 88


TthDNAPolymerase 89

TerminalDeoxynucleotidylTransferase,Recombinant(rTdT) 93

PolymixinBSulfate 235

PolyvinylPyrrolidone(PVP)—SeePVP-K-30 237

POMC 25

POMCGenePromoterReporterVector 111

PonceauS,SodiumSalt,Ultrapure 236

PotassiumChloride,Ultrapure 236

PotassiumChloride(KCl),2MSolution,Ultrapure 236

PotassiumPhosphate,Dibasic 236

PotassiumPhosphate,Dibasic,Ultrapure 236

PotassiumPhosphate,Monobasic 236

PotassiumPhosphate,Monobasic,Anhydrous,Ultrapure 236

PPARColdControlProbe 110

PPARaCMVExpressionVector 114

PPARaColdControlProbe 110

PPARaControlCos1NuclearExtract 110

PPARaELISAKit 110

PPARaELISAKitwithNuclearExtractionKit 110

PPARa-TFInteractionArrayI 118

PPARa-TFInteractionArrayII 118

PPARa-TFInteractionArrayIII 118

PPARb(PPARΔ)CMVExpressionVector 114



PPARg-TFInteractionArrayI 118

PPARg-TFInteractionArrayII 118

PPARg-TFInteractionArrayIII 118

PRLuciferaseReporterVector 112

PRA2 25

PREBCMVExpressionVector 114

Prep2SeqDNALibraryPrepKitforIllumina®platform 123,170

Prep2SeqMultiplexOligoAdaptersforIllumina®platform 124,171(Containsadaptersequences1-12)

Prep2SeqMultiplexOligoAdaptersforIllumina®platform 124,171(Containsadaptersequences13-27)

PrepEase8-wellStripStarterKitforVacuum 143

PrepEaseBACPurificationKit 145

PrepEaseDNACleanupKits 146

PrepEaseEndotoxin-FreeMaxiPlasmidKit 142

PrepEaseGelExtractionKits 146

PrepEaseGenomicDNAIsolationKits 147




PrepEaseGigaPlasmidKit 140

PrepEaseHistidine-TaggedHighSpecificityPurificationResin 155

PrepEaseHistidine-TaggedHighYieldPurificationResin 155

PrepEaseHistidine-TaggedProteinPurificationMaxiKit-HighSpecificity 153

PrepEaseHistidine-TaggedProteinPurificationMaxiKit-HighYield 154

PrepEaseHistidine-TaggedProteinPurificationMidiKit-HighSpecificity 153

PrepEaseHistidine-TaggedProteinPurificationMidiKit-HighYield 154

PrepEaseHistidine-TaggedProteinPurificationMiniKit-HighSpecificity 153

PrepEaseHistidine-TaggedProteinPurificationMiniKit-HighYield 154

PrepEaseMaxiPlasmidKit 140

PrepEaseMidiPlasmidKit 140

PrepEaseMiniSpinPlasmidKit 141

PrepEasemRNAMiniSpinKit 150

PrepEasePCRPurification96-WellPlateKits(Ultrafiltration) 148

PrepEasePlasmid8-wellStripKit 143

PrepEasePlasmid96-wellPlateCoreKit 144

PrepEasePlasmid96-wellPlateKit 144

PrepEaseProteinPurificationGlutathioneAgarose4B 155

PrepEaseQuickMiniSpinPlasmidKit 141

PrepEaseRNASpinKits 150

PrepEaseRNA/ProteinSpinKit 151

PrepEaseSequencingDyeCleanupKits 152

PrepEaseVacuumManifold 143

PrepEaseYeastPlasmidIsolationKit 145

PromoterMethylationPCRKit 24

ProteinAnalysis

AngiogenesisAntibodyArrays 133

CancerAntigenELISAKits 134

CytokineAntibodyArrays 134

ProteinMarkers

IEFProteinMarkers 134

ProteinMarkers,10-225kDa 134

ProteinMarkers,10-225kDa 136

Protein/DNAArrayI 116

Protein/DNAArrayIRefillKit 116

Protein/DNAArrayII 116

Protein/DNAArrayIIRefillKit 116

Protein/DNAArrayIII 116

Protein/DNAArrayIIIRefillKit 116

Protein/DNAArrayIV 116

Protein/DNAArrayIVRefillKit 116

Protein/DNAArrayV 116

Protein/DNAArrayVRefillKit 116

Proteinases

ProteinaseK 137

ProteinaseKSolution 137

ProteinaseK 137,237

ProteinaseKSolution 137

pUC19DNA—SeeMolecularBiologyProducts&Kitssection 129



PurificationSolutions

Phenol,pH8.0,Equilibrated 233

Phenol:Chloroform:IsoamylAlcohol(25:24:1) 233

PVP-K-30 237

Pyrophosphatase

Pyrophosphatase,Inorganic(Recombinant)(rPPase) 85

PyruvicAcid,SodiumSalt 237

R

D-(+)-Raffinose,Pentahydrate 237

RapidGel40%LiquidAcrylamideStockSolution,Ultrapure 237

RapidGel-XL6%LiquidAcrylamideDNASequencingGel,TBEFormulation,Ultrapure 238

RapidRunAgaroseBuffer,20XSolution,Ultrapure 238

RapidRunLoadingDye,Ultrapure 238

RARLuciferaseReporterVector 112

RB1GenePromoterReporterVector 111

Real-TimeqPCR

HotStart-ITProbeqPCRMasterMix(2X) 38

HotStart-ITProbeqPCRMasterMixwithUDG(2X) 38

HotStart-ITSYBRGreenqPCRMasterMix(2X) 32

HotStart-ITSYBRGreenqPCRMasterMixwithUDG(2X) 32

VeriQuestFastProbeqPCRMasterMix(2X) 36

VeriQuestFastProbeqPCRMasterMix,NoReferenceDye(2X) 37

VeriQuestFastSYBRGreenqPCRMasterMix(2X) 30

VeriQuestFastSYBRGreenqPCRMasterMixwithFluorescein 31

VeriQuestProbeqPCRMasterMix(2X) 33

VeriQuestProbeqPCRMasterMix,NoReferenceDye(2X) 35

VeriQuestSYBRGreenqPCRMasterMix(2X) 27

VeriQuestSYBRGreenqPCRMasterMixwithFluorescein(2X) 29

VeriQuestTaqDNAPolymerase 26




Real-TimeReverseTranscriptionPCR(qRT-PCR)


HotStart-ITProbeOne-StepqRT-PCRMasterMixKit 44

HotStart-ITSYBRGreenOne-StepqRT-PCRMasterMixKit 44

VeriQuestProbeOne-StepqRT-PCRMasterMix 42

VeriQuestProbeOne-StepqRT-PCRMasterMix,NoReferenceDye(2X) 43

VeriQuestSYBRGreenOne-StepqRT-PCRMasterMix(2X) 40

VeriQuestSYBRGreenOne-StepqRT-PCRMasterMixwithFluorescein(2X) 41

RecAProtein 65

ReporterStableCellLines

AP-1293ReporterStableCellLine 122

CHO/GFP-GR*StableCellLine 122

CHO/GFP-NFkBp65StableCellLine 122

CREB293ReporterStableCellLine 122

CREBCHOReporterStableCellLine 122

GR293ReporterStableCellLine 122

GRHeLaReporterStableCellLine 122

HIFNIH3T3ReporterStableCellLine 122

NFATHeLaReporterStableCellLine 122

NFATK562ReporterStableCellLine 122

NFkB293ReporterStableCellLine 122

NFkBA549ReporterStableCellLine 122

NFkBC2C12ReporterStableCellLine 122

NFkBHeLaReporterStableCellLine 122

NFkBNIH3T3ReporterStableCellLine 122

NFkBp50KnockdownHeLaStableCellLine 122

SRFHeLaReporterStableCellLine 122

STAT-1HeLaReporterStableCellLine 122

TAD-KinaseReporterStableCellLine 122

ReverseTranscriptases



ReverseTranscriptionRT-PCR-Kits


One-StepRT-PCRKit 47

Two-StepRT-PCRKit 48

ReverseTranscriptionRT-PCR-One-StepMasterMix

FideliTaqRT-PCRMasterMix(2X) 46

RT-PCRMasterMix(2X) 45

Riboflavin 238

D-(–)-Ribose 238

Rifampin 238

RNA,FreeAcid 238

RNA-ReverseTranscription



RNAPolymerases





RNaseInhibitors

RNaseInhibitor(HumanPlacenta) 94,164

RNaseInhibitor(Recombinant) 94,164

RNAPurification

PrepEasemRNAMiniSpinKit 150

PrepEaseRNA/ProteinSpinKit 151

PrepEaseRNASpinKits 150

RNATranscription





RNaseI 79,163

RNaseI‘A’,Powder 79,163

RNaseA,Lyophilized 79,163,239

RNaseA,MolecularBiologyGrade,Solution 80,163

RNaseH 80,163

RNaseInhibitor(HumanPlacenta) 94,164

RNaseInhibitor(Recombinant) 94,164

RNases

RNaseI 79

RnaseI'A'Powder 79

RNaseA,Lyophilized 79

RNaseA,MBGrade,Solution 80

RNaseH 80

ROXPassiveReferenceDye 53

RT-PCRMasterMix(2X) 45




RubyTaqDNAPolymerase 13

RubyTaqPCRMasterMix(2X) 14

RXRLuciferaseReporterVector 112

RXR(2)LuciferaseReporterVector 112

S

SAMe-PTS—SeeS-Adenosyl-L-MethionineSulfatep-Toluenesulfonate 190

SBECleanupReagent 52

SDS—SeeSodiumDodecylSulfate 240

SequenaseEnzyme&Kits

SequenasePCRProductSequencingKit 176

SequenaseQuickDenaturePlasmidDNASequencingKit 177


SequenaseVersion2.0DNASequencingKit 175

SequenasePCRProductSequencingKit 176

SequenaseQuickDenaturePlasmidDNASequencingKit 177

SequenaseRandomPrimerLabelingKit 127


SequenaseVersion2.0DNAPolymerasewithPyrophosphatase 85

SequenaseVersion2.0DNASequencingKit 175

SequencingDyeCleanup

PrepEaseSequencingDyeCleanupKits 152

DL-Serine(Non-Animal) 239



Single-StrandedDNABindingProtein(SSB) 65

SmadLuciferaseReporterVector 112

SMAD1(MADH1)CMVExpressionVector 114

SMAD-2ELISAKit 110

SMAD-2ELISAKitwithNuclearExtractionKit 110

Smad3/Smad4LuciferaseReporterVector 112

SMAD-4ELISAKit 110


SodiumAcetate(NaOAc),3MSolution,pH5.5,Ultrapure 239

SodiumAspartate—SeeAsparticAcid,SodiumSalt 201

SodiumAzide,Ultrapure 239

SodiumCacodylate—SeeCacodylicAcid,SodiumSalt 204

SodiumCarbonate,Anhydrous 239

SodiumCaseinate—SeeCasein,Sodium 205

SodiumChloride,Ultrapure 239

SodiumChloride(NaCl),5MSolution,Ultrapure 239

SodiumCitrate—SeeCitricAcid,TrisodiumSalt 207

SodiumDeoxycholate—SeeDeoxycholicAcid,SodiumSalt 210

SodiumDodecylSulfate(SDS) 240

SodiumDodecylSulfate(SDS),10%Solution,Ultrapure 240

SodiumDodecylSulfate(SDS),20%Solution,Ultrapure 240

SodiumDodecylSulfate(SDS),Ultrapure 240

SodiumGlutamate—SeeGlutamicAcid,MonosodiumSalt 218

SodiumLaurylSulfate—SeeSodiumDodecylSulfate 240

Sodium-N-LaurylSarcosine—SeeN-LaurylSarcosine,SodiumSalt 224

SodiumPhosphate,DibasicAnhydrous 240

SodiumPhosphate,Dibasic,Heptahydrate,Ultrapure 240

SodiumPhosphate,Monobasic 240

SodiumPhosphate,Monobasic,Monohydrate,Ultrapure 241

SodiumPyruvate—SeePyruvicAcid,SodiumSalt 237

SodiumSelenite 241

SolubleStarch—SeeStarch-Soluble 242

SodiumSuccinate—SeeSuccinicAcid,DisodiumSalt 242

D-Sorbitol,Ultrapure 241

SP-1ColdProbe 110

SP-1ControlRecombinantProtein 110

SP-1ELISAKit 110

SP-1ELISAKitwithNuclearExtractionKit 110

Sp1LuciferaseReporterVector 112

SP1(2)LuciferaseReporterVector 112

Spermidine,Trihydrochloride,Ultrapure 241

Spermine,Tetrahydrochloride 241

SRFHeLaReporterStableCellLine 122

SRELuciferaseReporterVector 112

SRFLuciferaseReporterVector 112

SRYCMVExpressionVector 114

SSC,20XSolution,Ultrapure 241

SSPE,20XSolution,Ultrapure 241

Starch,Hydrolyzed,Ultrapure 242

Starch-Soluble,Ultrapure 242

STAT-1ColdControlProbe 110

STAT-1ControlMCF7(IFNg) 110

STAT-1ELISAKit 110

STAT-1ELISAKitwithNuclearExtractionKit 110




STAT-1HeLaReporterStableCellLine 122

STAT1CMVExpressionVector 114

STAT1p84/p91LuciferaseReporterVector 112

STAT1GenePromoterReporterVector 111



STAT4LuciferaseReporterVector 112

STAT5BCMVExpressionVector 114

StockSolutions

AmmoniumAcetate(NH4OAc),5MSolution 199

Betaine,5MSolution 202

Dithiothreitol(DTT)0.1MSolution 213

EDTA,0.5MSolution 215

LithiumChloride(LiCl),7.5MSolution 225

MagnesiumChloride(MgCl2),1MSolution 226

PotassiumChloride(KCl),2MSolution 236

SodiumAcetate(NaOAc),3MSolution,pH5.5 239

SodiumChloride(NaCl),5MSolution 239

SodiumDodecylSulfate(SDS),10%Solution 240

SodiumDodecylSulfate(SDS),20%Solution 240

Water,Nuclease-Free 250

Water,RNase-Free,DEPCTreated 250

StopSolution,Ultrapure 242

Streptavidin,Ultrapure 242

Streptavidin-AlkalinePhosphataseConjugate 242

StreptomycinSulfate,Ultrapure 242

SuccinicAcid,DisodiumSalt,Hexahydrate 242

SuccinicAcid,FreeAcid 242

Sucrose 243

Sucrose,Crystalline 243

Sucrose,Ultrapure 243

SulfanilicAcid 243


Survivin 25

SURVIVINGenePromoterReporterVector 111

T

T3PromoterPrimer 178

T3RNAPolymerase,Highconcentration,200units/µl 92,160

T3RNAPolymerase,Standardconcentration,20units/µl 92,160

T4DNALigase,Highconcentration,10units/µl 70

T4DNALigase,Standardconcentration,1unit/µl 70


T4Gene32Protein,Highconcentration,≥10µg/µl 66

T4Gene32Protein,Standardconcentration,5µg/µl 66

T4PolynucleotideKinase(PNK) 69

T4RNALigase 71

T4RNALigase2 71

T7DNAPolymerase 88


T7RNAPolymerase,Highconcentration,>80units/µl 92,161

T7RNAPolymerase,Standardconcentration,20units/µl 92,161

TA-luc(EmptyControl)LuciferaseReporterVector 112

TAD-KinaseReporterStableCellLine 122

TAEBuffer,10XSolution,Ultrapure 243

TAEBuffer,50XSolution,Ultrapure 243

TaqDNAPolymerase 11

TaqPCRKit 12

TaqPCRMasterMix(2X) 12

Taurine,Ultrapure 243

TBEBuffer,5XSolution,Ultrapure 243

TBEBuffer,10XReady-MixedPowder,Ultrapure 243

TBS,20XSolution,pH7.4,Ultrapure 244

TBSwithTween(TBST),20XSolution,pH7.4,Ultrapure 244

TCEP-HCI,0.5MSolution,pH6.6±0.1 244





TEBuffer,1XSolution,Ultrapure 244

TEBuffer,50XSolution,Ultrapure 244

TEMED—SeeTetramethylethylenediamine 245

TerrificBroth,Ready-MadePowder 245

TerminalDeoxynucleotidylTransferase,Recombinant(rTdT) 93

TES,Ultrapure 245

Tetracycline,Hydrochloride,Ultrapure 245

TetramethylammoniumChloride,Ultrapure 245

3,3’,5,5’-Tetramethylbenzidine 245

N,N,N’,N’-Tetramethylethylenediamine,Ultrapure 245

TetrazoliumRed—See2,3,5-TriphenylTetrazoliumChloride 246




TFActivationELISAKits

AP-1ColdProbe 110

AP-1ELISAKit 110

AP-1ELISAKitwithNuclearExtractionKit 110

ControlHEK292PMANuclearExtract 110

E2F-1ELISAKit 110

E2F-1ELISAKitwithNuclearExtractionKit 110

ERaELISAKit 110

ERaELISAKitwithNuclearExtractionKit 110

EREColdProbe 110

ERControlMCF7NuclearExtract 110

FKHRELISAKit 110

FKHRELISAKitwithNuclearExtractionKit 110

GATA-1ColdProbe 110

GATA-1ControlRecombinantProtein 110

GATA-1ELISAKit 110

GATA-1ELISAKitwithNuclearExtractionKit 110

GRColdControlProbe 110

GRControlHela(DEX)NuclearExtract 110

GRELISAKit 110

GRELISAKitwithNuclearExtractionKit 110

HIFColdControlProbe 110

HIFControlCos1NuclearExtract 110

HIF-1aELISAKit 110

HIF-1aELISAKitwithNuclearExtractionKit 110

HNF-3bELISAKit 110

HNF-3bELISAKitwithNuclearExtractionKit 110

MEF-2ColdControlProbe 110

MEF-2ControlNuclearExtract 110

MEF-2ELISAKit 110

MEF-2ELISAKitwithNuclearExtractionKit 110

NFkBc-RelELISAKit 110

NFkBc-RelELISAKitwithNuclearExtractionKit 110

NFkBColdControlProbe 110

NFkBControlNuclearExtract 110

NFkBp50ELISAKit 110


NFkBp52ELISAKit 110


NFkBp65ELISAKit 110


NFkBRel-bELISAKit 110

NFkBRel-bELISAKitwithNuclearExtractionKit 110

p53ColdControlProbe 110

p53ELISAKit 110

p53ELISAKitwithNuclearExtractionKit 110

PPARColdControlProbe 110

PPARaColdControlProbe 110

PPARaControlCos1NuclearExtract 110

PPARaELISAKit 110

PPARaELISAKitwithNuclearExtractionKit 110

SMAD-2ELISAKit 110


SMAD-4ELISAKit 110


SP-1ColdProbe 110

SP-1ControlRecombinantProtein 110

SP-1ELISAKit 110

SP-1ELISAKitwithNuclearExtractionKit 110

STAT-1ColdControlProbe 110

STAT-1ControlMCF7(IFNg) 110

STAT-1ELISAKit 110

STAT-1ELISAKitwithNuclearExtractionKit 110

TF-TFInteractionArrayI 115

TF-TFInteractionArrayII 115

TF-TFInteractionArrayIII 115

TF-TFInteractionIRefill 115

TF-TFInteractionIIRefill 115

TF-TFInteractionIIIRefill 115

TFF1 25

THBS2GenePromoterReporterVector 111

ThermoSequenaseCycleSequencingKit 172

ThermoSequenaseDyePrimerManualCycleSequencingKit 173

ThermoSequenaseKits

ThermoSequenaseCycleSequencingKit 172

ThermoSequenaseDyePrimerManualCycleSequencingKit 173

Thiamine,Hydrochloride 245

ThiaminePyrophosphateChloride—SeeCocarboxylase 207




ThiazolylBlue—SeeMTT 228

Thimerosal 246

Thioredoxin,Human,Recombinant 246

L-Threonine 246

Thrombin 246

ThrombinHuman 246

TIMP3GenePromoterReporterVector 111

TMAC—SeeTetramethylammoniumChloride 245

TMB—See3,3’,5,5’-Tetramethylbenzidine 245

TNFaGenePromoterReporterVector 111

TopoisomeraseII,Alpha 95

ToxillicAcid—SeeMaleicAcid 226

TranscriptionFactors

EMSA(Gel-Shift)KitsandProbeSets 103

Function-SpecificProtein/DNAArrays 115

GenePromoterReporterVectors 111

LuciferaseReporterVectors 112

MammalianFull-lengthTFExpressionVectors 114

NFkB-TFandPPAR-TFInteractionArrays 118

Protein/DNAArrays 116

TFActivationELISAKits 107

TF-TFInteractionArrays 115

Transfection

DeliverXandDeliverXPlussiRNATransfectionKits 98

a,a-Trehalose,Dihydrate 246

Tricine,Ultrapure 246

Triethanolamine,Hydrochloride 246

1,3,7-Trimethylxanthine—SeeCaffeine 204

2,6,8-Trioxypurine—SeeUricAcid 249

2,3,5-TriphenylTetrazoliumChloride 246

TriphosphopyridineNucleotideDinucleotide—SeeNicotinamideAdenineDinucleotideReduced 230

2,4,6-Tripyridyl-s-Triazine(TPTZ) 247

Tris 247

Tris,Ultrapure 247

Tris-Glycine(TG)Buffer,10XSolution,Ultrapure 247

Tris-Glycine-SDS(TGS)Buffer,10XSolution,Ultrapure 247

Tris,Hydrochloride,Ultrapure 247

Tris/HCl,1MSolution,pH7.0,Ultrapure 248



r-hTRX—SeeThioredoxin,Human,Recombinant 246

Trypsin,Ultrapure 248

Trypsin1:250 248

Trypsin(TPCKTreated) 248

L-Tryptophan 248

TthDNAPolymerase 89

Two-StepRT-PCRKit 48

Tyrosinase 248

L-Tyrosine 248

U

Uracil 249

Uracil-DNAGlycosylase,E. coli 67

Uracil-DNAGlycosylase,Heat-Labile 67

Urea 249

Urea,Ultrapure 249

UricAcid 249

Uridine 249

Uridine-5’-Triphosphate,TrisodiumSalt 249

USF1CMVExpressionVector 114

UTP—SeeUridine-5’-Triphosphate 249

V

Vancomycin,Hydrochloride 250

VDRLuciferaseReporterVector 112

VeriQuestFastProbeqPCRMasterMix(2X) 36

VeriQuestFastProbeqPCRMasterMix,NoReferenceDye(2X) 37

VeriQuestFastSYBRGreenqPCRMasterMix(2X) 30

VeriQuestFastSYBRGreenqPCRMasterMixwithFluorescein(2X) 31

VeriQuestProbeOne-StepqRT-PCRMasterMix(2X) 42

VeriQuestProbeOne-StepqRT-PCRMasterMix,NoReferenceDye(2X) 43

VeriQuestProbeqPCRMasterMix(2X) 33

VeriQuestProbeqPCRMasterMix,NoReferenceDye(2X) 35

VeriQuestSYBRGreenOne-StepqRT-PCRMasterMix(2X) 40

VeriQuestSYBRGreenOne-StepqRT-PCRMasterMixwithFluorescein(2X) 41

VeriQuestSYBRGreenqPCRMasterMix(2X) 27

VeriQuestSYBRGreenqPCRMasterMixwithFluorescein(2X) 29

VeriQuestTaqDNAPolymerase 26

VHL 25




VHLGenePromoterReporterVector 111

VitafreeCasein—SeeCaseinVitafree 205

VitaminB1—SeeThiamineHCl 245

VitaminB2—SeeRiboflavin 238

VitaminB5—SeeD-CalciumPantothenate 204

VitaminB12 250

VitaminC—SeeL-AscorbicAcid 201

VitaminFreeCasein—SeeCasein-Vitafree 205

VitaminG—SeeRiboflavin 238

VitaminK1 250

v-mycGenePromoterReporterVector 111

W

Water,Nuclease-Free,Ultrapure 250

Water,RNase-Free,DEPCTreated,Ultrapure 250

Wright’sStain 250

X

XanthineMonosodiumSalt,Monohydrate 250

X-a-GAL—See5-Bromo-4-Chloro-3-Indolyl-a-D-Galactopyranoside 203

X-Gal—See5-Bromo-4-Chloro-3-Indolyl-b-D-Galactopyranoside 203

XyleneCyanolFF,Ultrapure 251

D-(+)-Xylose 251

Y, Z

Yeast,Bakers 251

Yeast,Brewers 251

YeastExtractPowder,LowDust 251

YeastExtractPowder,Ultrapure 251

YeastHydrolysate,Enzymatic,Ultrapure 251

Yeast-Torula 251

2XYTBroth 251

YY1LuciferaseReporterVector 112



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© 2014, 2015 Affymetrix, Inc. All rights reserved. CATALOG ISSUE 1.14FOR RESEARCH USE ONLY. Rev 10/15

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