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

Alexander Dobrovic, Tina Bianco, Lor Wai Tan, Tanya Sanders, Damian Hussey

Research output: Contribution to journalArticlepeer-review

19 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)134-138
Number of pages5
JournalMethods
Volume27
Issue number2
DOIs
Publication statusPublished - Jun 2002
Externally publishedYes

Keywords

  • Bisulfite
  • Cancer
  • Genomic sequencing
  • Methylation
  • p16
  • Silencing
  • Single-strand conformation analysis
  • Single-strand conformation polymorphism
  • Tumor

Fingerprint Dive into the research topics of 'Screening for and analysis of methylation differences using methylation-sensitive single-strand conformation analysis'. Together they form a unique fingerprint.

Cite this