Using native chromatin immunoprecipitation to interrogate histone variant protein deposition in embryonic stem cells

Zito Tseng, Tao Wu, Yifei Liu, Mei Zhong, Andrew Xiao

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Chromatin immunoprecipitation combined with massive parallel sequencing (ChIP-Seq) is a powerful epigenetics technique for interrogating the genome-wide localization of histone modifications, histone variants, and other chromatin-associating factors. In brief, chromatin pellets are fractionated from the nuclei, and then fragmented by enzymatic digestion or sonication. Chromatin regions associated with proteins of interest are enriched by immunoprecipitation with specific antibodies. After the immunoprecipitation, DNA fragments are extracted, amplified during sequencing library construction, and sequenced by high-throughput sequencing. Here, we describe the native chromatin immunoprecipitation of a rare histone variant, H2A.X, followed by high-throughput sequencing, in mouse embryonic stem cells.

Original languageEnglish (US)
Title of host publicationCancer Genomics and Proteomics
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages11-22
Number of pages12
ISBN (Print)9781493909919
DOIs
StatePublished - 2014

Publication series

NameMethods in Molecular Biology
Volume1176
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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