Urea improves efficiency of bisulphite-mediated sequencing of 5′-methylcytosine in genomic DNA

被引：103

作者：

Paulin, R

Grigg, GW

Davey, MW

Piper, AA

机构：

[1] Univ Technol Sydney, Dept Cell & Mol Biol, St Leonards, NSW 2065, Australia

[2] Peptech Ltd, Sydney, NSW 1670, Australia

来源：

NUCLEIC ACIDS RESEARCH | 1998年 / 26卷 / 21期

关键词：

D O I：

10.1093/nar/26.21.5009

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：

The detection of 5'-methylcytosine by the bisulphite-mediated genomic sequencing method has considerably aided study of the role of methylation in areas such as X chromosome inactivation, genomic imprinting and cancer research. However on occasion difficulty has been experienced in obtaining complete conversion of cytosine to uracil in regions of the target DNA. We report here a simple improvement to the method involving addition of urea to the bisulphite reaction, a step which greatly improves the reaction efficiency, presumably by maintaining the target DNA in single stranded form, thereby allowing complete and reliable conversion.

引用

页码：5009 / 5010

页数：2

共 8 条

[1]

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

[2] Demethylation of the human MDR1 5′ region accompanies activation of P-glycoprotein expression in a HL60 multidrug resistant subline [J].