Detection of DNA Methylation by MeDIP and MBDCap Assays: An Overview of Techniques

Methods Mol Biol. 2020:2102:225-234. doi: 10.1007/978-1-0716-0223-2_12.

Abstract

DNA methylation has been characterized as the representative example of epigenetic modifications and implicated in numerous biological processes, such as genomic imprinting and X chromosome inactivation. It primarily occurs at CpG dinucleotides in mammals and plays a critical role in transcriptional regulations. Examination of DNA methylation patterns in gene(s) or across a genome is vital to understand the role of epigenetic modulation in the progress of development and tumorigenesis. Currently, lots of approaches have been developed to investigate DNA methylation patterns for either limited regions or genome-scale studies, but some of them rely on using restriction enzymes. In this chapter, we describe two commonly used protocols to detect enrichment of methylated DNA regions, namely methylated immunoprecipitation (MeDIP) and capture of methylated DNA by methyl-CpG binding domain-based (MBD) proteins (MBDCap). They are the most economical and effective methods to study DNA methylation in either single locus or genome-wide scale.

Keywords: 5-Methylcytosine; CpG island; DNA methylation; MBDCap; MeDIP.

MeSH terms

  • 5-Methylcytosine / chemistry
  • Animals
  • Antibodies
  • CpG Islands / genetics
  • DNA / isolation & purification
  • DNA Methylation / genetics
  • DNA Restriction Enzymes
  • Epigenesis, Genetic
  • Epigenomics / methods*
  • Humans
  • Immunoprecipitation / methods*
  • Sonication / methods
  • Workflow

Substances

  • Antibodies
  • 5-Methylcytosine
  • DNA
  • DNA Restriction Enzymes