Screening for and analysis of methylation differences using methylation-sensitive single-strand conformation analysis

被引：17

作者：

Dobrovic, A

Bianco, T

Tan, LW

Sanders, T

Hussey, D

机构：

[1] Univ Adelaide, Dept Haematol Oncol, Adelaide, SA 5005, Australia

[2] Univ Adelaide, Dept Med, Adelaide, SA 5005, Australia

[3] Queen Elizabeth Hosp, Basil Hetzel Res Inst, Woodville, SA 5011, Australia

来源：

METHODS | 2002年 / 27卷 / 02期

基金：

英国医学研究理事会;

关键词：

methylation; cancer; tumor; silencing; bisulfite; single-strand conformation analysis; single-strand conformation polymorphism genomic sequencing; p16;

D O I：

10.1016/S1046-2023(02)00065-8

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Methylation-sensitive single-strand conformation analysis (MS-SSCA) is a method of screening for methylation changes at CpG sites in a region of DNA. After bisulfite modification. the region of interest is amplified using primers specific for bisulfite-modified sequences. The amplified products are denatured and run on a nondenaturing polyacrylamide gel. The sequence differences caused by methylation lead to the formation of different secondary structures (conformers) with different mobilities. MS-SSCA is a convenient and rapid method for screening large numbers of samples for methylation. Individual bands can readily be isolated and sequenced allowing more detailed analysis of methylation changes. In this article. we present a protocol for MS-SSCA and outline strategies for the design of primers for amplifying bisulfite-modified DNA sequences. (C) 2002 Elsevier Science (USA). All rights reserved,

引用

页码：134 / 138

页数：5

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