A BISULFITE METHOD OF 5-METHYLCYTOSINE MAPPING THAT MINIMIZES TEMPLATE DEGRADATION

被引：161

作者：

RAIZIS, AM ^{[1
]}

SCHMITT, F ^{[1
]}

JOST, JP ^{[1
]}

机构：

[1] FRIEDRICH MIESCHER INST,CH-4002 BASEL,SWITZERLAND

来源：

ANALYTICAL BIOCHEMISTRY | 1995年 / 226卷 / 01期

关键词：

D O I：

10.1006/abio.1995.1204

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The bisulfite method is a highly sensitive approach to 5-methylcytosine mapping that utilizes the capability of the polymerase chain reaction to exponentially amplify DNA. We have observed that the bisulfite reaction results in a significant level of template degradation due to DNA depurination. Furthermore, our data suggest that the DNA fragmentation which occurs limits the sensitivity of the method. We describe a simple solution to limit degradation of the DNA template. (C) 1995 Academic Press, Inc.

引用

收藏

页码：161 / 166

页数：6

相关论文

共 14 条

[1]

AMBROSE BJB, 1987, METHOD ENZYMOL, V152, P522

[2] GENOMIC SEQUENCING [J].

CHURCH, GM ;

GILBERT, W .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1984, 81 (07) :1991-1995

[3]

CLARK SJ, 1994, NUCLEIC ACIDS RES, V22, P2990, DOI 10.1093/nar/22.15.2990

[4] METHYLATION ANALYSIS ON INDIVIDUAL CHROMOSOMES - IMPROVED PROTOCOL FOR BISULFITE GENOMIC SEQUENCING [J].

FEIL, R ;

CHARLTON, J ;

BIRD, AP ;

WALTER, J ;

REIK, W .

NUCLEIC ACIDS RESEARCH, 1994, 22 (04) :695-696

[5] A GENOMIC SEQUENCING PROTOCOL THAT YIELDS A POSITIVE DISPLAY OF 5-METHYLCYTOSINE RESIDUES IN INDIVIDUAL DNA STRANDS [J].

FROMMER, M ;

MCDONALD, LE ;

MILLAR, DS ;

COLLIS, CM ;

WATT, F ;

GRIGG, GW ;

MOLLOY, PL ;

PAUL, CL .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (05) :1827-1831

[6]

Hayatsu H, 1976, Prog Nucleic Acid Res Mol Biol, V16, P75, DOI 10.1016/S0079-6603(08)60756-4

[7] TARGETED MUTATION OF THE DNA METHYLTRANSFERASE GENE RESULTS IN EMBRYONIC LETHALITY [J].

LI, E ;

BESTOR, TH ;

JAENISCH, R .

CELL, 1992, 69 (06) :915-926

[8] ROLE FOR DNA METHYLATION IN GENOMIC IMPRINTING [J].

LI, E ;

BEARD, C ;

JAENISCH, R .

NATURE, 1993, 366 (6453) :362-365

[9] A COMPUTER-PROGRAM FOR SELECTION OF OLIGONUCLEOTIDE PRIMERS FOR POLYMERASE CHAIN REACTIONS [J].

LOWE, T ;

SHAREFKIN, J ;

YANG, SQ ;

DIEFFENBACH, CW .

NUCLEIC ACIDS RESEARCH, 1990, 18 (07) :1757-1761

[10] PYRIMIDINE-SPECIFIC CHEMICAL-REACTIONS USEFUL FOR DNA SEQUENCING [J].

RUBIN, CM ;

SCHMID, CW .

NUCLEIC ACIDS RESEARCH, 1980, 8 (20) :4613-4619