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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

20 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

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

  1. SDG 3 - Good Health and Well-being
    SDG 3 Good Health and Well-being

Keywords

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

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