One-tube post-PCR fluorescent labeling of DNA fragments

被引：40

作者：

Inazuka, M ^{[1
]}

Tahira, T ^{[1
]}

Hayashi, K ^{[1
]}

机构：

[1] KYUSHU UNIV,INST GENET INFORMAT,DIV GENOME ANAL,HIGASHI KU,FUKUOKA 81282,JAPAN

来源：

GENOME RESEARCH | 1996年 / 6卷 / 06期

关键词：

D O I：

10.1101/gr.6.6.551

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 method for fluorescent postlabeling of PCR products has been developed. The method uses Klenow fragment of DNA polymerase I that exchanges the 3'-terminal residue of PCR-amplified DNA fragment for fluorescent nucleotides. All reactions, including PCR, are performed in one tube simply by successive addition of reagents. The products can be applied directly to fluorescence-based automated DNA sequencers without purification for either length determination in denaturing electrophoresis or mutation detection in SSCP electrophoresis.

引用

页码：551 / 557

页数：7

共 15 条

[1] CONTINUUM OF OVERLAPPING CLONES SPANNING THE ENTIRE HUMAN CHROMOSOME-21Q [J].

CHUMAKOV, I ;

RIGAULT, P ;

GUILLOU, S ;

OUGEN, P ;

BILLAUT, A ;

GUASCONI, G ;

GERVY, P ;

LEGALL, I ;

SOULARUE, P ;

GRINAS, L ;

BOUGUELERET, L ;

BELLANNECHANTELOT, C ;

LACROIX, B ;

BARILLOT, E ;

GESNOUIN, P ;

POOK, S ;

VAYSSEIX, G ;

FRELAT, G ;

SCHMITZ, A ;

SAMBUCY, JL ;

BOSCH, A ;

ESTIVILL, X ;

WEISSENBACH, J ;

VIGNAL, A ;

RIETHMAN, H ;

COX, D ;

PATTERSON, D ;

GARDINER, K ;

HATTORI, M ;

SAKAKI, Y ;

ICHIKAWA, H ;

OHKI, M ;

LEPASLIER, D ;

HEILIG, R ;

ANTONARAKIS, S ;

COHEN, D .

NATURE, 1992, 359 (6394) :380-387

[2] NOVEL NON-TEMPLATED NUCLEOTIDE ADDITION-REACTIONS CATALYZED BY PROCARYOTIC AND EUKARYOTIC DNA-POLYMERASES [J].

CLARK, JM .

NUCLEIC ACIDS RESEARCH, 1988, 16 (20) :9677-9686

[3] HOW SENSITIVE IS PCR-SSCP [J].

HAYASHI, K ;

YANDELL, DW .

HUMAN MUTATION, 1993, 2 (05) :338-346

[4] SINGLE-STRAND CONFORMATIONAL POLYMORPHISM USING CAPILLARY ELECTROPHORESIS WITH 2-DYE LASER-INDUCED FLUORESCENCE DETECTION [J].

HEBENBROCK, K ;

WILLIAMS, PM ;

KARGER, BL .

ELECTROPHORESIS, 1995, 16 (08) :1429-1436

[5]

IWAHANA H, 1995, PCR METH APPL, V4, P275

[6] DNA SEQUENCING WITH DYE-LABELED TERMINATORS AND T7 DNA-POLYMERASE - EFFECT OF DYES AND DNTPS ON INCORPORATION OF DYE-TERMINATORS AND PROBABILITY ANALYSIS OF TERMINATION FRAGMENTS [J].

LEE, LG ;

CONNELL, CR ;

WOO, SL ;

CHENG, RD ;

MCARDLE, BF ;

FULLER, CW ;

HALLORAN, ND ;

WILSON, RK .

NUCLEIC ACIDS RESEARCH, 1992, 20 (10) :2471-2483

[7]

Makino R, 1992, PCR Methods Appl, V2, P10

[8]

MASHIYAMA S, 1990, Technique (Philadelphia), V2, P304

[9] RAPID AND SENSITIVE DETECTION OF POINT MUTATIONS AND DNA POLYMORPHISMS USING THE POLYMERASE CHAIN-REACTION [J].

ORITA, M ;

SUZUKI, Y ;

SEKIYA, T ;

HAYASHI, K .

GENOMICS, 1989, 5 (04) :874-879

[10] ANALYSIS OF THE CD3 GENE REGION AND TYPE-1 DIABETES - APPLICATION OF FLUORESCENCE-BASED TECHNOLOGY TO LINKAGE DISEQUILIBRIUM MAPPING [J].

PRITCHARD, LE ;

KAWAGUCHI, Y ;

REED, PW ;

COPEMAN, JB ;

DAVIES, JL ;

BARNETT, AH ;

BAIN, SC ;

TODD, JA .

HUMAN MOLECULAR GENETICS, 1995, 4 (02) :197-202

← 1 2 →