Characterization of in vivo DNA-binding events of plant transcription factors by ChIP-seq: Experimental protocol and computational analysis

Hilda Van Mourik*, J.M. Muiño, Alice Pajoro, G.C. Angenent, Kerstin Kaufmann

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

14 Citations (Scopus)

Abstract

Chromatin immunoprecipitation followed by next-generation sequencing (ChIP-seq) is a powerful technique for genome-wide identification of in vivo binding sites of DNA-binding proteins. The technique had been used to study many DNA-binding proteins in a broad variety of species. The basis of the ChIP-seq technique is the ability to covalently cross-link DNA and proteins that are located in very close proximity. This allows the use of an antibody against the (tagged) protein of interest to specifically enrich DNAfragments bound by this protein. ChIP-seq can be performed using antibodies against the native protein or against tagged proteins. Using a specific antibody against a tag to immunoprecipitate tagged proteins eliminates the need for a specific antibody against the native protein and allows more experimental flexibility. In this chapter we present a complete workflow for experimental procedure and bioinformatic analysis that allows wet-lab biologists to perform and analyze ChIP-seq experiments.

Original languageEnglish
Title of host publicationPlant Functional Genomics: Methods and Protocols: Second Edition
PublisherSpringer
Pages93-121
ISBN (Print)9781493924448, 9781493924431
DOIs
Publication statusPublished - 2015

Keywords

  • ChIP-seq data analysis
  • Chromatin immunoprecipitation
  • Plant transcription factors
  • Transcription factor DNA-binding sites

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