Methylation-sensitive, single-strand conformation analysis (MS-SSCA): A rapid method to screen for and analyze methylation

Tina Bianco, Damian Hussey, Alexander Dobrovic

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

We have developed methylation, sensitive, single-strand conformation analysis (MS-SSCA) as a method of screening for methylation changes. Bisulfite modification converts cytosines to thymines, but methylated cytosines remain unchanged. This modification creates sequence differences between methylated and unmethylated samples, which can be resolved by SSCA. SSCA is 70-95% efficient at detecting single base changes in a fragment. As bisulfite modification of methylated DNA would typically involve several base changes in a fragment, the efficiency of detecting methylation using MS-SSCA could approach 100%. We applied this method to analyze the BRCA1 promoter CpG island in breast cancer samples. About 20% of sporadic breast cancers are hypermethylated at the BRCA1 promoter CpG island. MS-SSCA rapidly detected those tumors that had previously been shown to be methylated by Southern blotting. The variant bands detected by SSCA were analyzed by sequencing and shown to be methylated. MS-SSCA is a simple method for screening large numbers of samples for methylation and can accelerate genomic sequencing, as all bands can be isolated and sequenced directly.

Original languageEnglish
Pages (from-to)289-293
Number of pages5
JournalHuman Mutation
Volume14
Issue number4
DOIs
Publication statusPublished - Oct 1999
Externally publishedYes

Keywords

  • BRCA1
  • Cancer
  • Detection
  • Methylation
  • MS-SSCA
  • Single-strand conformation analysis

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