Methylation-sensitive single-molecule analysis of chromatin structure

Curr Protoc Mol Biol. 2010 Jan:Chapter 21:Unit 21.17.1-16. doi: 10.1002/0471142727.mb2117s89.

Abstract

Methylation-sensitive single-molecule analysis of chromatin structure is a high-resolution method for studying nucleosome positioning. As described in this unit, this method allows for the analysis of the chromatin structure of unmethylated CpG islands or in vitro-remodeled nucleosomes by treatment with the CpG-specific DNA methyltransferase SssI (M.SssI), followed by bisulfite sequencing of individual progeny DNA molecules. Unlike nuclease-based approaches, this method allows each molecule to be viewed as an individual entity instead of an average population.

MeSH terms

  • Animals
  • Biocatalysis
  • Chromatin / chemistry*
  • CpG Islands
  • DNA Methylation*
  • Genetic Techniques*
  • Humans

Substances

  • Chromatin