amplification pcr reagents - korambiotech.com · our expertise in enzyme engineering has produced...

PCR Reagents

+ How do you ensure optimal amplification results?

+ How do you choose the best PCR enzyme for your application?

+ How can you improve cDNA synthesis?

AMPLIFICATION

PCR REAGENTS 0>0

We have the best PCR reagent for your application.

Our expertise in enzyme engineering has produced one of the broadest portfolios of

high-fidelity PCR enzymes available today. This expertise has translated to advances in

high-fidelity reverse transcription PCR. Whether you need the highest accuracy

available, large product yields, or sensitive cDNA synthesis, trust our high-performance

enzymes to answer your amplification challenges.

PCR Enzymes & Master Mixes

6 Ultra-High Fidelity PCR

7 High Fidelity for TA/UA Cloning

8 High Fidelity for Blunt-End Cloning

9 High Sensitivity & High Yield

10 GC-Rich/Complex Targets

11 Long PCR

12 Routine PCR

+

CONTENTS:

RT-PCR Kits & Reagents

13 RT-PCR

PCR-Related Kits & Reagents

14 Nucleotides

14 PCR Purification Kits

14 PCR Cloning Kits

15 Ordering Information

www.stratagene.com

PCR ENZYMES

Easy-A™

High Fidelity PCR

oning Enzym

Easy-A™

High Fidelity PCR

Master Mix

PfuTurbo®o®

Hotstart DNA

Polymerase

PfuTurbo®®

DNA Polymerase

PfuTurbo®o®

Hotstart PCR

Master Mix

PfuUltra™a™

Hotstart DNA

Polymerase

PfuUltra™a™

High-Fidelity

DNA Polymerase

PfuUltra™a™

Hotstart PCR

Master Mix

High Fidelity (Contamination

Control)

Ultra-High Fidelity (Cloning &

Mutagenesis)

PfuTurbo®o®CxCxx

Hotstart DNA

Polymerase

High Fidelity(TA/UA Cloning)

High Fidelity (Blunt Cloning)

Format

RT-PCR KITS & REAGENTS

StrataScript™

One-Tube RT-PCR Kit with

Easy-A™ PCR Cloning Enzyme

StrataScript™

Two-Tube RT-PCR Kit with

PfuUltra™ DNA Polymerase

StrataScript™

First Strand

Synthesis Kit

PicoMaxx™ PCR System

HighFidelity

High Yield/Sensitivity

Format

Herculase®

Hotstart DNA

Polymerase

Herculase®

nhanced DN

Polymerase

Herculase®

Hotstart PCR

Master Mix

EXL™

DNA Polymerase

PicoMaxx™

High Fidelity

PCR System

PicoMaxx™

High Fidelity

PCR Master Mix

Taq2000 ™

DNA Polymerase

High Sensitivity/High Yield

Long PCR Routine PCRGC-Rich/Complex

TemplatesFormat

Hotstart Enzyme

Standard Enzyme

Master Mix

Hotstart Enzyme

Standard Enzyme

Master Mix

cDNA Synthesis

One-Tube RT-PCR

Two-Tube RT-P

PCR REAGENTS4>5

The Importance of High Fidelity High-fidelity PCR enzymes are valuable for minimizing the introduction of amplification

errors in products that will be cloned, sequenced, and expressed. Significant time and

effort can be saved by employing high-fidelity amplification procedures that eliminate the

need for downstream error-correction steps and minimize the number of clones that must

be sequenced in order to obtain error-free constructs or accurate consensus sequences.

Moreover, the use of high-fidelity amplification conditions is essential when analyzing

small amounts of template DNA or rare molecules in heterogeneous populations.

Amplifications employing small amounts of template DNA are especially prone to high

mutant frequencies due to PCR-generated errors in early cycles1, 2.

The ArchaeMaxx® Factor AdvantageBecause proofreading PCR enzymes can be finicky, we have improved high-fidelity PCR to

be more reliable. Our ArchaeMaxx® Polymerase-Enhancing Factor, found exclusively in our

high-fidelity DNA polymerases, improves overall PCR performance by eliminating

the inhibition of proofreading enzymes caused by incorporation of dUTP, which results

from deamination of dCTP during PCR 3. Our PCR enzymes formulated with the

ArchaeMaxx factor produce higher yields and exhibit greater target length capability.

Superior Reverse TranscriptionOur expertise in enzyme engineering also extends to reverse transcriptases. Both MMLV

and AMV reverse transcriptases possess substantial RNase H activity that degrades

RNA molecules and limits full-length cDNA synthesis and yield. For this reason, RTs that

lack RNase H activity are far superior to MMLV and other RNAse H+ RTs. Our StrataScript™

Reverse Transcriptase contains a point mutation that effectively abolishes RNase H activi-

ty, allowing more efficient production of full-length cDNA than RNase H+ RTs.

www.stratagene.com

Comparison of Our PCR Enzymes

StrataScript™ First Strand Synthesis Kit 0-9 kb + +

StrataScript™ First-Strand Synthesis Kit 0-9 kb + + +

plus PicoMaxx™ High Fidelity PCR System

StrataScript™ One-Tube RT-PCR Kit 0-5 kb + +

with Easy-A™ PCR Cloning Enzyme

StrataScript™ Reverse Transcriptase 0-9 kb + +

StrataScript™ Two-Tube Kit 0-9 kb + + +

with PfuUltra™ DNA Polymerase

RT-PCR Reagent TargetLength

RNase H Deficient

cDNA Synthesis

One-Tube RT-PCR

Two-Tube RT-PCR

Comparison of Our RT-PCR Kits & Reagents

PCR Enzyme Target Length

Blunt or 3'-A Ends

ArchaeMaxx®

AdvantageHot StartAvailable

Master MixAvailable

Accuracy vs. Taq

PfuUltra™ High Fidelity DNA Polymerase 18x0-17 kb (genomic)

Blunt + + +0-15 kb (vector)

PfuTurbo® DNA Polymerase 6x 0-19 kb (genomic)Blunt + + +

0-15 kb (vector)

PfuTurbo® Cx DNA Polymerase 6x 0-10 kb Blunt +

Pfu DNA Polymerase, cloned or native 6x 0-5 kb Blunt

Easy-A™ High Fidelity PCR Cloning Enzyme 6x 0-5 kb 3'-A + + +

Herculase® DNA Polymerase 3x 0-37 kb Mixed + + +

EXL™ DNA Polymerase 3x20-37 kb (genomic)

Mixed +20-50 kb (vector)

PicoMaxx™ High Fidelity PCR System 2x 0-10 kb Mixed + + +

SureStart® Taq DNA Polymerase 1x 0-5 kb 3'-A +

Taq2000 ™ DNA Polymerase 1x 0-4 kb 3'-A

Our continuing efforts to improve the fidelity of PCR have resulted in PfuUltra™ High-FidelityDNA Polymerase, the most accurate PCR enzyme commercially available. PfuUltra DNA poly-merase is ideal for PCR cloning, site-directed mutagenesis, and anywhere sequence accuracy is critical for downstream applications.

The Most Accurate PCR Enzyme

Highest Fidelity Available

The PfuUltra™ high-fidelity DNA polymerase ø,‡ contains a

genetically engineered mutant of Pfu DNA polymerase***,‡ that

delivers 300% greater accuracy. A validated and referenced

fidelity assay4 demonstrates that PfuUltra high-fidelity DNA poly-

merase exhibits an average error rate three-fold lower than Pfu

DNA polymerase and 18-fold lower than Taq DNA polymerase,

making it the most accurate PCR enzyme available (Figure 1).

Enhanced Performance

In addition to ultra-high fidelity, PfuUltra high-fidelity DNA

polymerase exhibits superior PCR performance compared to

most PCR proofreading enzymes. The addition of our ArchaeMaxx

polymerase-enhancing factor promotes higher yields, shorter

extension times, and greater target length capability (Figure 2).

High Specificity Hotstart Version

The PfuUltra™ Hotstart DNA Polymeraseø,‡ is an antibody-mediated

hotstart formulation of PfuUltra high fidelity DNA polymerase

that provides higher specificity and reduced background.

Convenient Master Mix Formulation

For ultra-high fidelity with added convenience and increased

throughput, choose PfuUltra™ Hotstart PCR Master Mix ø,‡.

This 2x formulation contains PfuUltra hotstart DNA polymerase,

buffer, magnesium, and dNTPs all in one tube for faster setup

and more consistent results.

Ultra-High Fidelity PCR

PCR REAGENTS > PCR ENZYMES & MASTER MIXES6>7

0 5 10 15 20 25

PfuUltra™ DNA Polymerase

PfuTurbo™ and PfuTurbo® Cx DNA Polymerases

Tgo DNA Polymerase

Herculase® DNA Polymerase

Pfx/KOD DNA Polymerase

Expand™ High Fidelity

Advantage-HF™ Polymerase

Taq DNA Polymerase

ACCURACY x 105DNA PolymerasesM 0.97 1.5 3.5 kb171296

Figure 1THE MOST ACCURATE PCR ENZYME AVAILABLE PfuUltra™ High-Fidelity DNA Polymerase provides the greatest accuracy of any commercially

available enzyme1. Fidelity was measured using our validated and referenced fidelity assay4.

Accuracy = 1/Error Rate.

Figure 2AMPLIFIES A WIDE RANGE OF TARGETSPfuUltra™ High-Fidelity DNA Polymerase amplifies genomic

DNA targets up to 17 kb.

PCR Enzyme Accuracy vs. Taq

Blunt or 3'-A Ends

ArchaeMaxx®


Master MixAvailable

PfuUltra™

High Fidelity DNA Polymerase

18x Blunt + + +

TargetLength

0-17 kb (genomic)0-15 kb (vector)

High Fidelity for TA/UA Cloning

TA or UA cloning requires PCR products with 3'-A overhangs. We created the Easy-A™ High-Fidelity PCR CloningEnzyme to automatically add A-overhangs for quick and easy high-fidelity cloning into TA/UA vectors.

Unique Proofreading Formulation Adds A-Overhangs

Combines Cloning Efficiency with High Accuracy

The Easy-A™ high-fidelity PCR cloning enzyme ,‡,† is the only

proofreading DNA polymerase formulation to deliver high-

throughput TA or UA cloning. PCR products amplified with the

Easy-A PCR cloning enzyme can be cloned directly to T- or

U-modified vectors with high cloning efficiency and throughput

(Figure 3). But unlike Taq DNA polymerase, the Easy-A PCR

cloning enzyme exhibits proofreading activity that provides the

accuracy level of Pfu DNA polymerase.

Fidelity Without Extra Work

Adapting blunt-ended fragments amplified with proofreading

enzymes requires post-PCR addition of 3'-A overhangs prior

to the cloning step. This additional step includes a secondary

incubation with Taq DNA polymerase and requires the opening

and closing of tubes, which exposes the lab to contamination

risk. The Easy-A high-fidelity PCR cloning enzyme does not

require any post-PCR A-addition steps.

High Specificity Hotstart Formulation

The Easy-A high fidelity PCR cloning enzyme is provided as

an antibody-mediated hotstart formulation, providing higher

specificity and reduced background.


For the highest throughput in TA or UA cloning, choose Easy-A™

High-Fidelity PCR Master Mix ,‡,†. This 2x formulation contains the

Easy-A PCR cloning enzyme, buffer, magnesium, and dNTPs all

in one tube for faster setup and more consistent results.

www.stratagene.com

Figure 3HIGH CLONING EFFICIENCY WITH EASY-A™ PCR CLONING ENZYMETargets amplified with the Easy-A™ High-Fidelity PCR Cloning Enzyme contain 3'-A overhangs, allowing quick and easy

cloning into any T- or U-modified vector.

T-vectorT-vector

Ligation complete

Topoisomerase-mediated ligationor conventional ligation

5' 3'

3'

5'

3'

5'

3'

5'

PCRProduct

TT

TTAA

AAEasy-A-Amplified

PCR Products

Add 0.5-1.5 µl of PCR product to T-modified vector

T-vector T-vector

U-vectorU-vector

Ligation complete

Ligation

5' 3'

3'

5'

3'

5'

3'

5'

PCRProduct

UU

UUA

A

AAEasy-A-Amplified

PCR Products

Add 0.5-1.5 µl of PCR product to U-modified vector

U-vector U-vector

™™


Blunt or 3'-A Ends

ArchaeMaxx®

AdvantageHot StartFormat

Master MixAvailable

Easy-A™ HighFidelity PCRCloning Enzyme

6x 3'-A + + +

TargetLength

0-5 kb

We invented the high-fidelity PCR market over a decade ago with the original Pfu DNA Polymerase.Since that time, our expertise has led to improvements in high-fidelity performance with PfuTurbo®

DNA Polymerase and, more recently, PfuTurbo® Cx Hotstart DNA Polymerase. These improvementshave made high-fidelity PCR more reliable and versatile.

Robust High-Fidelity Amplification

The PfuTurbo® DNA polymerase ø,‡ is a special formulation of

cloned Pfu DNA polymerase and our ArchaeMaxx polymerase-

enhancing factor that produces increased PCR product yields

without affecting fidelity. The enhanced performance of PfuTurbo

DNA polymerase allows the use of shorter extension times,

longer targets, and lower concentrations of DNA template than

are suitable for Pfu DNA polymerase.

High Fidelity for Blunt-End Cloning


9 kb

6 kb

2.6 kb

0.9 kb

PFUTURBO ® Cx ACCUPRIME® PFXM PLATINUM® PFX

Figure 4SUPERIOR PERFORMANCE WITH PFUTURBO® CX HOTSTART DNA POLYMERASEAmplification of a range of targets from 0.9 to 9 kb using PfuTurbo® Cx Hotstart DNA

Polymerase and other commercially available proofreading PCR enzymes.


The PfuTurbo® hotstart DNA polymerase ø,‡ is an antibody-

mediated hotstart formulation of PfuTurbo DNA polymerase that

provides higher specificity, reduced background, and enhanced

yield of challenging systems.

New Mutant Overcomes Uracil Sensitivity

Heat can cause deamination of cytosine to uracil in a DNA

strand. To prevent replication of such a mutation, archaeal

(proofreading) polymerases typically stop replication just before

reaching a uracil residue in the template5. In PCR, this poisoning

phenomenon can result in decreased or no product yield.

We formulated PfuTurbo ® Cx hotstart DNA polymerase**,‡ with

a novel mutant of Pfu DNA polymerase that can read through

a uracil located in the template strand without stalling. Thus

PfuTurbo Cx hotstart DNA polymerase improves the overall

reliability of high-fidelity PCR and exhibits less finicky, more

robust performance (Figure 4).

Prevent Carryover Contamination

Because PfuTurbo Cx hotstart DNA polymerase can replicate

uracil-containing DNA, it can be used to prevent carry-over

contamination. This method involves the use of dUTP in place

of dTTP in the nucleotide pool. Subsequent PCR reactions are

pre-treated with Uracil-N-glycosylase (UNG) to degrade any

dU-containing DNA left over from a previous reaction; the UNG

is then inactivated during the next denaturation step. PfuTurbo Cx

hotstart DNA polymerase incorporates dUTP in targets up to 6 kb

in length more efficiently than Taq DNA polymerase.

Accuracy with Enhanced Performance


Blunt or 3'-A Ends

ArchaeMaxx®


Master MixAvailable

PfuTurbo ®

DNA Polymerase6x Blunt + + +

TargetLength


PfuTurbo ® CxDNA Polymerase 6x Blunt +0-10 kb

Pfu DNAPolymerase,cloned or native

6x Blunt 0-5 kb

High Sensitivity & High Yield

Blending Taq DNA polymerase with a proofreading enzyme improves target-length capability, fidelity,and yield, but many PCR enzyme blends lack sensitivity and can lead to PCR failures. We designed thePicoMaxx™ High Fidelity PCR System to deliver the highest sensitivity and efficiency, bringing yougreater blend performance at a much lower cost per unit than other enzyme blends.

Greater Performance at a Fraction of the Cost

Sensitivity and Reliability

The PicoMaxx™ high fidelity PCR system ø,‡ is designed to provide

maximum PCR sensitivity and reliability. Sensitivity is critical for

successful amplification, especially with limited or precious sam-

ples; use of a PCR enzyme that lacks sensitivity often results in

amplification failures. Formulated with a blend of Taq and Pfu

DNA polymerases and our ArchaeMaxx polymerase-enhancing

factor, together with a specially optimized buffer, the PicoMaxx

high fidelity PCR system overcomes such PCR failures with

greater sensitivity than other PCR enzymes (Figure 5). Superior

sensitivity makes the PicoMaxx high fidelity PCR system one of

the most reliable PCR enzyme formulations available.

High Efficiency and Yield

The PicoMaxx high fidelity PCR system exhibits superior amplifi-

cation efficiencies due in large part to our patented ArchaeMaxx

factor, which eliminates dUTP that accumulates during the high

heat of PCR and can poison high-fidelity reactions.

The ArchaeMaxx factor maximizes the performance of the proof-

reading component of the blend, allowing the PicoMaxx high

fidelity PCR system to deliver higher product yields and more

reliable performance across a wide range of targets up to 10 kb.


For maximum sensitivity and yield with added convenience,

choose PicoMaxx™ High Fidelity PCR Master Mix ø,‡. This 2x

formulation contains the PicoMaxx high fidelity enzyme blend,

buffer, magnesium, and dNTPs all in one tube for faster setup

and more consistent results.

www.stratagene.com

Figure 5PICOMAXX™ HIGH FIDELITY PCR SYSTEM EXHIBITS SUPERIOR SENSITIVITYWe amplified a 3.9 kb α-1 anti-trypsin template with PicoMaxx™ High Fidelity PCR System, Expand™

High Fidelity PCR System (Roche), and Platinum® Taq DNA Polymerase High Fidelity (Invitrogen).

We performed all reactions according to the manufacturer’s recommendations.

PICOMAXX™

HIGH FIDELITYPLATINUM® TAQHIGH FIDELITY

EXPAND™

HIGH FIDELITYGenomic DNATemplate Qty.

(ng)

3.9 kb

50 25 10 1 50 25 10 1 50 25 10 1


Blunt or 3'-A Ends

ArchaeMaxx®


Master MixAvailable

PicoMaxx™

High Fidelity PCR System

2x Mixed + + +

TargetLength

0-10 kb

Some PCR targets are problematic to amplify due to their content or structure. Our Herculase®

Enhanced DNA Polymerase helps overcome these PCR challenges with successful amplification of complex and GC-rich templates over a wide range of template sizes.

Overcome Challenging PCR


The Herculase® Hotstart DNA Polymerase ø,‡ is an antibody-

mediated hotstart formulation of Herculase enhanced DNA

polymerase that provides higher specificity, reduced back-

ground, and enhanced yield of challenging systems.


For added convenience and increased throughput, choose

Herculase® Hotstart PCR Master Mixø,‡. This 2x formulation

contains Herculase hotstart DNA polymerase, buffer,

magnesium, and dNTPs all in one tube for faster setup and

more consistent results.

GC-Rich/Complex Targets


COMPETITOR’S GC-RICH PCR ENZYME+/- GC-Rich Solution (M)

HERCULASE®

+/- DMSO (%)

0 06 7 8 9 0.5 1.0 1.5 2.0

1 2 3 4 5 6 7 8 9 10M

Figure 6HERCULASE® ENHANCED DNA POLYMERASE EXCELS IN AMPLIFYING GC-RICH TARGETS Herculase® Enhanced DNA Polymerase easily amplifies an 82.5% GC-rich fragment of the Fragile X

gene from human genomic DNA.


Blunt or 3'-A Ends

ArchaeMaxx®


Master MixAvailable

Herculase®

DNA Polymerase 3x Mixed + + +

TargetLength

0-37 kb

GC-Rich and Complex Templates

The Herculase® enhanced DNA polymeraseø,‡ meets today’s

complex PCR challenges. When you need to accurately amplify

a complex or GC-rich template, Herculase DNA polymerase will

help you succeed. A unique optimized formulation of high fidelity

Pfu DNA polymerase, Taq DNA polymerase and our ArchaeMaxx

polymerase-enhancing factor, Herculase DNA polymerase excels

in amplifying a broad range of target lengths with superior fidelity

than Taq DNA polymerase and other PCR enzyme blends6.

DMSO is provided separately as a PCR adjunct, and can be

added when amplifying difficult targets to increase product yield

and extend target-length capability (Figure 6).

Long PCR

To amplify very long targets, a blend of Taq DNA polymerase and a proofreader provides the bestresults. But not every enzyme blend can go the distance. Our EXL™ DNA Polymerase is speciallyoptimized to successfully and accurately amplify long targets up to 50 kb.

Go the Full Distance

Tackle Long Amplicons

The EXL™ DNA polymerase ø,‡ provides superior performance

in amplifying complex targets greater than 20 kb in length.

The EXL DNA polymerase is a special formulation of Pfu DNA

polymerase, Taq DNA polymerase, and our ArchaeMaxx

polymerase enhancing factor, along with a special buffer

optimized specifically for very long targets. This enzyme

blend provides robust yields and reliable amplification (Figure 7).

Amplify with Higher Accuracy

Because PCR enzymes have an intrinsic rate at which they

incorporate incorrect bases, long templates are particularly

prone to errors when amplified during PCR. Most “Long &

Accurate” PCR enzyme blends available commercially provide

only a slight increase in accuracy over Taq. EXL DNA poly-

merase provides twice the accuracy of these other blends

to ensure minimal errors in your long PCR products.

www.stratagene.com

Figure 7EXL™ DNA POLYMERASE EXCELS AT AMPLIFYING EXTREMELY LONG TARGETSEXL™ DNA Polymerase easily amplifies two extremely long genomic targets (23 and 30 kb)

and a lambda target (45 kb). A competitor’s PCR enzyme for long targets is shown for

comparison. All reactions were performed according to the manufacturer’s recommendations.

EXL™

M 23 30 45

COMPETITOR

23 30 45


Blunt or 3'-A Ends

ArchaeMaxx®

AdvantageHot Start Available

Master MixAvailable

EXL™ DNAPolymerase 3x Mixed +

TargetLength


When fidelity is not of concern, our Taq2000™ DNA Polymerase provides consistentamplification of simple targets. For applications requiring higher specificity, includingreal-time QPCR, choose SureStart® Taq DNA Polymerase for hotstart capability.

High-Quality Taq Enzymes Provide Consistent Results

Consistent Performance

The Taq2000 ™ DNA polymerase‡ is a highly purified, recombinant

Taq DNA polymerase cloned from the thermophilic eubacterium,

Thermus aquaticus. Using Taq2000 DNA polymerase in longer

PCR amplifications reduces smearing and virtually eliminates

unwanted background artifacts. Taq2000 DNA has superior

thermostability compared to other commercial Taq DNA

polymerase preparations.

Hotstart for Maximum Specificity

Our SureStart® Taq DNA polymerase‡ is a hot-start Taq DNA

polymerase that can be incorporated into PCR protocols previously

optimized with Taq DNA polymerase, with little modification of

cycling parameters or reaction conditions. This specially modified

Taq DNA polymerase allows setup of PCR reactions at ambient

room temperature without nonspecific annealing and extension

during PCR setup, making setup easier. SureStart Taq DNA

polymerase can be used in a variety of amplification systems

to improve specificity, yield, and amplification of difficult targets.

Efficient, Sensitive, Real-Time PCR Quantification

As part of our rigorous quality control testing, every lot of

SureStart Taq DNA polymerase is routinely validated for QPCR.

For maximum performance in real-time PCR, choose one of our

Brilliant® QPCR and QRT-PCR reagent kits or master mixes‡, α,

each of which is formulated with SureStart Taq DNA polymerase.

Routine PCR



Blunt or 3'-A Ends

ArchaeMaxx®


Master MixAvailable

SureStart® TaqDNA Polymerase

1x 3'-A +

TargetLength

0-5 kb

Taq2000 ™

DNA Polymerase 1x 3'-A0-14 kb

RT-PCR

Most common reverse transcriptases, including MMLV and AMV, exhibit RNase H activity,which limits yields of full-length cDNA. With StrataScript™ Reverse Transcriptase we have eliminated RNase H activity to provide greater yields of full-length transcripts.

Produce High cDNA Yields

Mutant RT Delivers Higher Yields and Sensitivity

The StrataScript™ reverse transcriptase is a Moloney murine

leukemia virus reverse transcriptase (MMLV RT) that has

been genetically modified to remove RNase H activity without

affecting the desired reverse transcriptase function. As a result,

StrataScript RT generates longer transcripts and greater

performance. We manufacture StrataScript RT to be free from

impurities, allowing for the highest possible cDNA yield (Figure 8).

StrataScript RT also provides superior sensitivity compared to

MMLV and AMV reverse transcriptases in real-time QRT-PCR

applications. StrataScript RT is the ideal choice to generate

cDNA for a variety of downstream applications including library

construction and RT-PCR.

High-Quality First-Strand Synthesis

The StrataScript™ First Strand cDNA Synthesis Kit§ employs

StrataScript RT to generate cDNA of high quality from total or

poly(A) RNA. Subsequent amplification with sequence-specific

primers yields a homogenous population of the specific cDNA

molecule of interest.

One-Tube RT-PCR System for Cloning

The StrataScript™ One-Tube RT-PCR System‡ combines high

cloning efficiency with high fidelity in a one-tube RT-PCR format.

The kit features our robust StrataScript RT and our Easy-A PCR

cloning enzyme, which amplifies with six-fold higher accuracy

thanTaq DNA polymerase yet adds 3'-A overhangs for efficient

TA or UA cloning.

Ultra-High Fidelity RT-PCR

The StrataScript™ Two-Tube RT-PCR System‡ is the highest

fidelity RT-PCR system available. Our StrataScript RT is

included for highly efficient first strand synthesis. The cDNA

is then amplified with PfuUltra high fidelity DNA polymerase.

The StrataScript two-tube RT-PCR system is ideal for

applications where sequence integrity is critical.

High Sensitivity Two-Tube RT-PCR

When sensitivity is important, and you want to archive your

cDNA, combine our StrataScript first strand cDNA synthesis kit

with our PicoMaxx™ high fidelity PCR system. The PicoMaxx

PCR system provides the most sensitive amplification of any

PCR enzyme or blend available.

www.stratagene.com

Figure 8SAME ROBUST PERFORMANCEReactions using 100 U of StrataScript™ Reverse Transcriptase and 100 U of SuperScript™ II

Reverse Transcriptase (Invitrogen) generated equivalent performance at 40ºC.

100 USUPERSCRIPT™ II

RT

100 USTRATASCRIPT™

RT

RT-PCR REAGENTS > RT-PCR KITS & REAGENTS

RT-PCRReagent

TargetLength

cDNASynthesis

One-TubeRT-PCR

Two-TubeRT-PCR

StrataScript™ One-Tube RT-PCR Kitwith Easy-A™ PCR Cloning Enzyme

0-5 kb +

+

RNase HDeficient

+

StrataScript™ Two-Tube Kit withPfuUltra™ DNA Polymerase 0-9 kb +

+StrataScript™ First Strand Synthesis Kit

0-9 kb +

+

We offer a wide range of kits and reagents to complete your PCR and prepare for a variety of downstream applications.

High-Quality PCR-Grade Deoxynucleotide Mix

Our dNTP mix contains 400 µl of 100 mM dNTP mix (25 mM

each of dATP, dCTP, dGTP, and dTTP), adequate for 1000-2000

standard primer-extension reactions. This mix produces high-

quality reaction products and withstands multiple freeze-thaw

cycles without compromising efficiency.

Safe, Easy PCR Cleanup

Our StrataPrep® PCR Purification Kits prepare PCR products

quickly and easily for downstream applications such as

sequencing, microarray analysis, and cloning. Following

amplification or cDNA labeling, a DNA binding solution is added

directly to the PCR tube and then transferred to a unique

microspin cup. Once bound, the DNA (cDNA) is washed and

then eluted from the column, ready for resuspension in your

application buffer of choice. For high-throughput applications,

our StrataPrep® 96 PCR Purification Kit provides the same easy

and efficient cleanup in a convenient 96-well format.

Efficient PCR Cloning

PCR-Script® Cloning Kits§, allow the efficient cloning of PCR

fragments with a high yield and a low rate of false positives.

PCR products are incubated with one of the predigested

PCR-Script cloning vectors, Srf I and T4 DNA ligase. Using

the restriction enzyme in the ligation reaction maintains a high

steady-state concentration of digested vector DNA and allows the

use of nonphosphorylated, unmodified PCR primers. The ligation

efficiency of blunt-ended DNA fragments is increased by the

simultaneous, opposite reactions of the Srf I restriction enzyme

and T4 DNA ligase on nonrecombinant vector DNA. The PCR-

Script cloning kits include the StrataPrep PCR purification kit

and also include polishing reagents to create blunt-ended PCR

products amplified with a non-proofreading enzyme.

PCR-Related Kits & Reagents

PCR-RELATED KITS & REAGENTS14

Ordering Information

Product Name Size CatalogUltra-High Fidelity

PFUULTRA™ HIGH FIDELITY DNA POLYMERASE 100 U 600380

500 U 600382

1000 U 600384

PFUULTRA™ HOTSTART DNA POLYMERASE 100 U 600390

500 U 600392

1000 U 600394High Fidelity, TA or UA Cloning

EASY-A™ HIGH FIDELITY PCR CLONING ENZYME 100 U 600400

500 U 600402

1000 U 600404High Fidelity, Blunt Cloning

PFU DNA POLYMERASE, CLONED 100 U 600153

500 U 600154

1000 U 600159

PFU DNA POLYMERASE, NATIVE 100 U 600135

500 U 600136

1000 U 600140

PFUTURBO® CX HOTSTART DNA POLYMERASE 100 U 600410

500 U 600412

1000 U 600414

PFUTURBO® DNA POLYMERASE 100 U 600250

500 U 600252

1000 U 600254

PFUTURBO® HOTSTART DNA POLYMERASE 100 U 600320

500 U 600322

1000 U 600324High Sensitivity & Yield

PICOMAXX™ HIGH FIDELITY PCR SYSTEM 100 U 600420

500 U 600422

1000 U 600424GC-Rich/Complex Targets

HERCULASE® ENHANCED DNA POLYMERASE 100 U 600260

500 U 600262

1000 U 600264

HERCULASE® HOTSTART DNA POLYMERASE 100 U 600310

500 U 600312

1000 U 600314Extra-Long Targets

EXL™ DNA POLYMERASE 100 U 600340

500 U 600342

1000 U 600344Routine PCR

SURESTART® TAQ DNA POLYMERASE 100 U 600280

500 U 600282

1000 U 600284

TAQ2000™ DNA POLYMERASE 100 U 600195

500 U 600196

1000 U 600197

www.stratagene.com

PCR Enzymes PCR Master Mixes

LEGAL

* U.S. Patent Nos. 6,734,293, 6,444,428, 6,379,553, 6,333,165, 6,183,997 and patents pending.

** U.S. Patent Nos. 6,734,293, 6,444,428, 6,379,553, 6,333,165, 6,183,997, 6,489,150, 5,948,663, 5,866,395,5,556,772, 5,545,552 and patents pending.

*** U.S. Patent Nos. 6,489,150, 5,948,663, 5,866,395, 5,545,552 and patents pending.

Patents pending

ø U.S. Patent Nos. 6,734,293, 6,489,150, 6,444,428, 6,379,553, 6,333,165, 6,183,997, 5,948,663, 5,866,395, 5,556,772, 5,545,552 and patents pending.

‡ Purchase of these products is accompanied by a license to use them in the Polymerase Chain Reaction (PCR) process in conjunction with a thermal cycler whose use in the automated performance of the PCR process is covered by the up-front license fee, either by payment to Applied Biosystems or as purchased, i.e., an authorized thermal cycler.

§ Purchase of this PCR-related product does not convey any rights under the PCR patents owned by Roche MolecularSystems. A license to use the PCR process accompanies the purchase of certain reagents from Stratagene when used in conjunction with an Authorized Thermal Cycler.

† Use of these products for certain applications may require licenses from third parties in certain countries.

Use of the CMV promoter is covered under U.S. Patent Nos. 5,168,062 and 5,385,839 owned by the University of Iowa Research Foundation and licensed FOR RESEARCH USE ONLY.

α Use of labeling reagents may require licenses from entities other than Stratagene. For example, use of fluorogenic probesin 5' nuclease assays may require licenses under U.S. Patent Nos. 6,214,979, 5,804,375, 5,210,015 and 5,487,972 ownedby Roche Molecular Systems, Inc. and under U.S. Patent No. 5,538,848 owned by Applied Biosystems.

Expand is a trademark of Roche Diagnostics Corporation.

Platinum is a registered trademark of Invitrogen Corporation.

SuperScript is a trademark of Invitrogen Corporation.

REFERENCES

1. Hogrefe, H.H. and M.C. Borns. High Fidelity PCR Enzymes. In C.W. Dieffenbach, G.S. Dveksler (eds.) PCR Primer: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 2003.

2. Cha, R.S. and W.G. Thilly. Specificity, efficiency, and fidelity of PCR. In PCR Primer: A Laboratory Manual (eds, Dieffenbach, C.W. and G.S. Dveksler) Cold Spring Harbor Laboratory Press, 1995.

3. Hogrefe, et al. (2002). Proc. Nat. Acad. Sci.USA 99: 596-601.

4. Cline, J., Braman, J.C. and Hogrefe, H.H. (1996) Nucleic Acids Res. 24: 3546-3551.

5. Fogg, et al. (2002). Nat Struct Biol. 9(12): 922-927.

6. Borns, M. and Hogrefe, H. (2000) Strategies. 13: 12.

Product Name Size CatalogUltra-High Fidelity

PFUULTRA™ HOTSTART PCR MASTER MIX 100 rxn 600630

400 rxn 600632High Fidelity, TA or UA Cloning

EASY-A™ HIGH FIDELITY PCR MASTER MIX 100 rxn 600640

400 rxn 600642High Fidelity, Blunt Cloning

PFUTURBO® HOTSTART PCR MASTER MIX 100 rxn 600600

400 rxn 600602High Sensitivity & Yield

PICOMAXX™ HIGH FIDELITY PCR MASTER MIX 100 rxn 600650GC-Rich/Complex Targets

HERCULASE® HOTSTART PCR MASTER MIX 100 rxn 600610

400 rxn 600612

RT-PCR Reagents

RT-PCR

STRATASCRIPT™ FIRST STRAND SYNTHESIS KIT 50 rxn 200420

STRATASCRIPT™ ONE-TUBE RT-PCR KIT 50 rxn 600168WITH EASY-A™ PCR CLONING ENZYME

STRATASCRIPT™ REVERSE TRANSCRIPTASE, 50U/µL 10,000U 600085

STRATASCRIPT™ REVERSE TRANSCRIPTASE, 200U/µL 10,000U 600086

STRATASCRIPT™ TWO-TUBE RT-PCR KIT 50 rxn 600170WITH PFUULTRA™ DNA POLYMERASE

PCR-Related Kits & Accessories

Nucleotide Mix

DEOXYNUCLEOTIDE MIX, PCR-GRADE (G/A/T/C, 25 mM EACH) 400 µl 200415PCR Purification

STRATAPREP® 96 PCR PURIFICATION KIT 2 plates 400775

10 plates 400776

50 plates 400774

STRATAPREP® PCR PURIFICATION KIT 50 preps 400771

250 preps 400773PCR Cloning Kits

PCR-SCRIPT® AMP CLONING KIT 10 rxn 211188

25 rxn 211190

50 rxn 211189

PCR-SCRIPT® CAM CLONING KIT 25 rxn 211192

ORDERING INFORMATION

AMPLIFICATION CELL BIOLOGY CLONING MICROARRAYS NUCLEIC ACIDANALYSIS

PROTEIN FUNCTION& ANALYSIS

QUANTITATIVEPCR

SOFTWARESOLUTIONS

Stratagene USA and Canada

Order: 800-424-5444 x3Technical Services: 800-894-1304

Stratagene Europe

Order: 00800-7000-7000Technical Services: 00800-7400-7400

Stratagene Japan K.K.

Order: 03-5159-2060Technical Services: 03-5159-2070

Distributors >For a list of worldwide distributors, please visit:

www.stratagene.com

11011 North Torrey Pines RoadLa Jolla, CA 92037 USA

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