Whole-genome DNA methylation profiling using MethylCap-seq

Methods. 2010 Nov;52(3):232-6. doi: 10.1016/j.ymeth.2010.06.012. Epub 2010 Jun 11.

Abstract

MethylCap-seq is a robust procedure for genome-wide profiling of DNA methylation. The approach consists of the capture of methylated DNA using the MBD domain of MeCP2, and subsequent next-generation sequencing of eluted DNA. Elution of the captured methylated DNA is done in steps using a salt gradient, which stratifies the genome into fractions with different CpG density. The enrichment reached within the individual eluates allows for cost-effective deep sequence coverage. The profiles together yield a detailed genome-wide map of methylated regions and readily allows detection of DNA methylation in known and novel regions. Here, we describe principles and details of the MethylCap-seq procedure using different sources of starting material.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • DNA / chemistry
  • DNA / metabolism*
  • DNA Methylation*
  • Genome, Human / genetics*
  • Humans
  • Methyl-CpG-Binding Protein 2 / metabolism*
  • Polymerase Chain Reaction / methods
  • Sequence Analysis, DNA / methods

Substances

  • Methyl-CpG-Binding Protein 2
  • DNA