Chromatin immunoprecipitation-based analysis of gene regulatory networks operative in human embryonic stem cells

Methods Mol Biol. 2012:873:269-80. doi: 10.1007/978-1-61779-794-1_18.

Abstract

Chromatin immunoprecipitation (ChIP) followed by microarray-based (ChIP-Chip) or next-generation sequencing-based (ChIP-Seq) analysis has been established as a powerful and widely used method to investigate DNA-protein interactions relative to a genomic location in vivo. Here, we present a ChIP-Chip protocol, which utilizes an alternative, easier amplification protocol and when using high-quality ChIP-grade antibodies, will generate enough material for hybridization or sequencing with negligible enrichment bias due to amplification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatin / genetics
  • Chromatin / metabolism
  • Chromatin Immunoprecipitation / methods*
  • Embryonic Stem Cells / metabolism*
  • Gene Regulatory Networks / genetics
  • Gene Regulatory Networks / physiology*
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Humans
  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3 / genetics
  • Octamer Transcription Factor-3 / metabolism
  • Oligonucleotide Array Sequence Analysis

Substances

  • Chromatin
  • Homeodomain Proteins
  • NANOG protein, human
  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3
  • POU5F1 protein, human