Enrichment of methylated DNA by methyl-CpG immunoprecipitation

Methods Mol Biol. 2013:971:201-12. doi: 10.1007/978-1-62703-269-8_11.

Abstract

Normal DNA methylation is an epigenetic modification required for proper development. Aberrant DNA methylation, in contrast, is frequently observed in many different malignancies including leukemias and lymphomas. Global DNA methylation profiling addresses the methylated sequences (methylome) of patient genomes to identify disease-specific methylation patterns. Workload in methylome analyses can be considerably reduced by methylome enrichment using proteins or antibodies with high affinity to methylated DNA. Methyl-CpG Immunoprecipitation (MCIp) employs an immobilized recombinant human methyl-CpG binding domain protein 2, MBD2, which binds methylated CpGs in double-stranded DNA. Elution with increasing salt concentrations allows the fractionated enrichment of different degrees of methylation.

MeSH terms

  • CpG Islands*
  • DNA / genetics
  • DNA Methylation*
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Genomics
  • Humans
  • Immobilized Proteins / chemistry
  • Immobilized Proteins / genetics
  • Immobilized Proteins / metabolism
  • Immunoprecipitation / methods*
  • Magnets / chemistry
  • Microspheres
  • Sonication
  • Staphylococcal Protein A / chemistry

Substances

  • DNA-Binding Proteins
  • Immobilized Proteins
  • MBD2 protein
  • Staphylococcal Protein A
  • DNA