Direct sequencing of unpurified PCR-amplified DNA by semi-exponential cycle sequencing (SECS)

被引：5

作者：

Sarkar, G ^{[1
]}

Bolander, ME ^{[1
]}

机构：

[1] Mayo Clin, Rochester, MN 55905 USA

来源：

MOLECULAR BIOTECHNOLOGY | 1997年 / 8卷 / 03期

关键词：

DNA sequencing; PCR amplification;

D O I：

10.1007/BF02760780

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A simple technique for direct sequencing of PCR-amplified templates without purification of the PCR reaction product is presented. This method does not require an additional synthesis step after template amplification, and can generate sequence information form as little as 0.1 fmol of unpurified template.

引用

收藏

页码：269 / 277

页数：9

相关论文

共 11 条

[1] DNA SEQUENCING WITH THERMUS-AQUATICUS DNA-POLYMERASE AND DIRECT SEQUENCING OF POLYMERASE CHAIN REACTION-AMPLIFIED DNA [J].

INNIS, MA ;

MYAMBO, KB ;

GELFAND, DH ;

BROW, MAD .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (24) :9436-9440

[2] RAPID SEQUENCING OF UNPURIFIED PCR PRODUCTS BY THERMAL ASYMMETRIC PCR CYCLE SEQUENCING USING UNLABELED SEQUENCING PRIMERS [J].

LIU, YG ;

MITSUKAWA, N ;

WHITTIER, RF .

NUCLEIC ACIDS RESEARCH, 1993, 21 (14) :3333-3334

[3]

MELTZER SJ, 1990, BIOTECHNIQUES, V8, P142

[4] A SYSTEM FOR SHOTGUN DNA SEQUENCING [J].

MESSING, J ;

CREA, R ;

SEEBURG, PH .

NUCLEIC ACIDS RESEARCH, 1981, 9 (02) :309-321

[5]

MULLIS KB, 1987, METHOD ENZYMOL, V155, P335

[6] A RAPID POLYMERASE-CHAIN-REACTION-DIRECTED SEQUENCING STRATEGY USING A THERMOSTABLE DNA-POLYMERASE FROM THERMUS-FLAVUS [J].

RAO, VB ;

SAUNDERS, NB .

GENE, 1992, 113 (01) :17-23

[7]

SANDHU GS, 1989, BIOTECHNIQUES, V7, P689

[8] DNA SEQUENCING WITH CHAIN-TERMINATING INHIBITORS [J].

SANGER, F ;

NICKLEN, S ;

COULSON, AR .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1977, 74 (12) :5463-5467

[9]

Sarkar G, 1993, PCR Methods Appl, V2, P318

[10] SEMI EXPONENTIAL CYCLE SEQUENCING [J].

SARKAR, G ;

BOLANDER, ME .

NUCLEIC ACIDS RESEARCH, 1995, 23 (07) :1269-1270