Chromatin Immunoprecipitation

Methods Mol Biol. 2018:1839:65-75. doi: 10.1007/978-1-4939-8685-9_7.

Abstract

Chromatin immunoprecipitation (ChIP) measures the physical association between a protein and DNA in the cell. In combination with next-generation sequencing, the technique enables the identification of DNA targets for the corresponding protein across an entire genome. Here we describe the immunoprecipitation of Vibrio cholerae DNA bound to the histone-like nucleoid structuring protein (H-NS) tagged with the Flag epitope. The quality of the DNA obtained in this protocol is suitable for next-generation sequencing. The procedure described herein can be readily adapted to other bacteria and DNA-binding proteins.

Keywords: Chromatin immunoprecipitation; DNA-binding proteins; Immunoprecipitation; Next-generation sequencing; Transcription; Vibrio cholerae.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Bacterial Proteins / metabolism
  • Chromatin Immunoprecipitation*
  • DNA-Binding Proteins
  • High-Throughput Nucleotide Sequencing
  • Protein Binding
  • Real-Time Polymerase Chain Reaction
  • Vibrio cholerae / genetics
  • Vibrio cholerae / metabolism

Substances

  • Bacterial Proteins
  • DNA-Binding Proteins