Multiplexed ChIP-Seq Using Direct Nucleosome Barcoding: A Tool for High-Throughput Chromatin Analysis

Methods Mol Biol. 2018:1689:177-194. doi: 10.1007/978-1-4939-7380-4_16.

Abstract

Chromatin immunoprecipitation followed by sequencing (ChIP-Seq) or microarray hybridization (ChIP-on-chip) are standard methods for the study of transcription factor binding sites and histone chemical modifications. However, these approaches only allow profiling of a single factor or protein modification at a time.In this chapter, we present Bar-ChIP, a higher throughput version of ChIP-Seq that relies on the direct ligation of molecular barcodes to chromatin fragments. Bar-ChIP enables the concurrent profiling of multiple DNA-protein interactions and is therefore amenable to experimental scale-up, without the need for any robotic instrumentation.

Keywords: ChIP-Seq; Chromatin; Immunoprecipitation; MNase; Multiplex; NGS; Nucleosomes; Saccharomyces cerevisiae.

MeSH terms

  • Chromatin / genetics
  • Chromatin / metabolism
  • Chromatin Immunoprecipitation* / methods
  • Computational Biology / methods
  • DNA Barcoding, Taxonomic*
  • Data Interpretation, Statistical
  • Gene Library
  • High-Throughput Nucleotide Sequencing* / methods
  • Nucleosomes / genetics*
  • Nucleosomes / metabolism
  • Real-Time Polymerase Chain Reaction / methods
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Workflow

Substances

  • Chromatin
  • Nucleosomes