Chromatin Immunoprecipitation for Analyzing Transcription Factor Binding and Histone Modifications in Drosophila

Methods Mol Biol. 2016:1478:263-277. doi: 10.1007/978-1-4939-6371-3_16.

Abstract

Chromatin immunoprecipitation followed by next-generation sequencing (ChIP-seq) is an invaluable technique to assess transcription factor binding and histone modifications in a genome-wide manner, an essential step towards understanding the mechanisms that govern embryonic development. Here, we provide a detailed protocol for all steps involved in generating a ChIP-seq library, starting from embryo collection, fixation, chromatin preparation, immunoprecipitation, and finally library preparation. The protocol is optimized for Drosophila embryos, but can be easily adapted for any model organism. The resulting library is suitable for sequencing on an Illumina HiSeq or MiSeq platform.

Keywords: ChIP-seq; Chromatin immunoprecipitation; Development; Drosophila; Next-generation sequencing; Transcription.

Publication types

  • Review

MeSH terms

  • Animals
  • Chromatin / chemistry
  • Chromatin / metabolism
  • Chromatin Assembly and Disassembly
  • Chromatin Immunoprecipitation / methods
  • Drosophila melanogaster / genetics*
  • Drosophila melanogaster / growth & development
  • Drosophila melanogaster / metabolism
  • Embryo, Nonmammalian
  • Gene Library
  • Genome*
  • High-Throughput Nucleotide Sequencing
  • Histones / genetics*
  • Histones / metabolism
  • Protein Binding
  • Protein Processing, Post-Translational*
  • Signal Transduction
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Transcription, Genetic*

Substances

  • Chromatin
  • Histones
  • Transcription Factors