Whole-genome DNA methylation profiling using MethylCap-seq.
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.
Brinkman
Arie B
AB
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
Simmer
Femke
F
Ma
Kelong
K
Kaan
Anita
A
Zhu
Jingde
J
Stunnenberg
Hendrik G
HG
D045744:Cell Line, Tumor; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D019175:DNA Methylation; D015894:Genome, Human / Q000235:genetics*; D006801:Humans; D051783:Methyl-CpG-Binding Protein 2 / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA / Q000379:methods
D051783:Methyl-CpG-Binding Protein 2; D004247:DNA