DNA extraction from serum and plasma

Features

By using DNA Extractor SP Kit, DNA was extracted from 10 human serum and one plasma samples. The extracted DNA was finally resolved in 20μL of TE (pH8.0), and 5μL of that was subject to amplification of p53-Exon 5 (308bp). Commercial available kit based on a glass binding method (spin column method) was used as a comparative method.

３０８bp

PCR ： 40 cycles 3% Agarose gel

M ： DNA Step Ladder Mix（80ｰ10kbp）

W ： DNA Extractor SP Kit

A ： Competitor's kit

Application data 1 : Result of amplification of p53-Exon 5 region extracted from human serum and plasma

M W A W A W A W A W A W A W A W A W A W A W A

Serum 1 Serum 2 Serum 3 Serum 4 Plasma Serum 5 Serum 6 Serum 7 Serum 8 Serum 9 Serum 10

■High DNA yield (about 100%) from small amount of serum and plasmaThe DNA extraction procedure is based on a simple centrifugal separation using Sodium Iodide and alcohol, that results in less loss of DNA.

■Safe operationNo phenol or chloroform required

■Minimum contamination The entire extraction of DNA can be done in a single centrifuge tube.

■Complete removal of lipid derived from bloodA specialized alcohol solution in original formula

■Less variability in extraction from sample to sample

DNA Extractor SP Kit efficiently extracts DNA fragments contained in serum and plasma. Based on the Sodium Iodide (NaI) method1, this kit enables the whole procedures done in a single micro-centrifuge tube without using hazardous phenol/chloroform.Recently, tumor-specific genes have been amplified and detected in the serum and plasma of patients with various diseases such as lung, breast and colon cancer. Many of these reference articles are currently being published.The kit is a powerful pretreatment reagent for the detection and analysis of target DNA because of its high quality and yield.

2,3,4

Wako Catalog No. 296-60501 50 tests

Keep at 2～10℃

for Genetic Research

Catalog No.Product name Usable samples Package Size

Plasma, serum 50 testsDNA Extractor SP Kit 296-60501

【Product List】

【Related Products】

【References】

【Storage condition】

1）Ishizawa, M., Kobayashi, Y., Miyamura, T. and Matsuura, S. : Nucleic Acids Res., 22, 1774（1994）.2）Sozzi, G., Musso, K., Ratcliffe, C., Goldstraw, P., Pierotti, M. A. and Pastorino, U.: Clin Cancer Res., 5, 2689（1999）.3）Silva, J. M., Dominguez, G., Garcia, J. M., Gonzalez, R., Villanueva, M. J., Navarro, F., Provencio, M., San, Martin, S., Espana, P. and Bonilla,　F.: Cancer Res., 59 , 3251（1999）.4）Shao, Z. M., Wu, J., Shen, Z. Z. and Nguyen, M.: Clin Cancer Res. 7, 2222（2001）.

４３９bp

Serum 2 and 3 are those used in the application data 1

PCR : 40 cycles 3% Agarose gel

M ： DNA Step Ladder Mix 　　 （80ｰ10kbp） NC ： Negative control (no DNA) W ： DNA Extractor SP Kit A ： Competitor's kit

Kit Contents : 50 reactions ●Enzyme reaction solution 1×10mL ●Alcohol solution 1×30mL●Protein digestion solution 1×250μL ●Washing Solution（A） 1×50mL●Sodium iodide solution 1×15mL ●Washing Solution（B） 1×50mL

100μL sample of serum or plasma＋ 200μL of Enzyme Reaction Solution and mix

briefly.＋ 5μL of Protein Digestion Solution and vortex. Incubate at 56℃ for 10 min. ＋ 300μL of Sodium Iodide Solution and mix

briefly. ＋ 600μL of alcohol solution and vortex. Incubate at room temperature for 10 min.

Centrifuge at 12,000～20,000×g for 10 min. at room temperature.

Pellet＋ 1mL of Washing Solution (A) to the pellet

and vortex.Centrifuge at 12,000～20,000×g for 5 min. at room temperature.

Pellet＋ 1mL of Washing Solution (B) to the pellet

and vortex. Centrifuge at 12,000～20,000×g for 5 min. at room temperature.

Pellet Dry the pellet well for about 5 min. at about 65°C. ＋ Adequate volume (10～20μL) of TE (pH 8.0)

or D.W. and vortex. Dissolve the pellet completely with vortex mixer and incubation at about 65°C for 3～5 min.

DNA solution

Standard protocol

Application data 2 : Result of amplification of p53-Exon 7 region extracted from human serum

Catalog No.Product name Usable samples Package size

Serum, biopharmaceuticals

Whole blood

Whole blood

Hair, bloodstain

Human

Human

50 tests

50 tests

20 tests

200 tests

50 tests

100 tests

100 tests

DNA Extractor Kit

DNA Extractor WB Kit

DNA Extractor WB-Rapid Kit

DNA Extractor FM Kit

P53 Primer Exon 5

P53 Primer Exon 7

295-50201

291-50502

297-54801

293-54803

295-58501

312-03511

316-03531

DNA was extracted from Serum 2 and 3 used in the test data 1 and another human serum (Serum 11) for amplification of p53-Exon 7 (439bp).

M W A W A W A W A

NC Serum 3 Serum 2 Serum 11

DNA Extractor SP Kit

Wako Pure Chemical Industries, Ltd.http://www.wako-chem.co.jp1-2, Doshomachi 3-ChomeChuo-ku, Osaka 540-8605, JapanTel: 81-6-6203-3741Fax: 81-6-6201-5964Online Cat:http://search.wako-chem.com

Wako Chemicals USA, Inc.http://www.wakousa.comHead Office:1600 Bellwood Road, Richmond, VA 23237Toll-Free (U.S. only): 1-877-714-1920Tel: 1-804-714-1920/ Fax: 1-804-271-7791Los Angeles Sales Office:15625 Alton Parkway, Suite D, Irvine, CA 92618Tel: 1-949-679-1700/ Fax: 1-949-679-1701

Wako Chemicals GmbHhttp://www.wako-chemicals.deEuropean Office:Nissanstraße 2, D-41468Neuss, GermanyTel: 49-2131-311-0Fax: 49-2131-311100

05303IBH

Keep at 2～10℃