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

Whole-genome DNA methylation profiling using MethylCap-seq.

Author information

1
Radboud University, Department of Molecular Biology, Nijmegen Centre for Molecular Life Sciences, Faculty of Science, 6500 HB Nijmegen, The Netherlands. a.brinkman@ncmls.ru.nl

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.

PMID:
20542119
DOI:
10.1016/j.ymeth.2010.06.012
[Indexed for MEDLINE]

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