A cell-sorting-based protocol for cell cycle small-scale ChIP sequencing

Johnathan R. Whetstine, Capucine Van Rechem

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Classic approaches to characterizing cell cycle leverage chemicals or altered nucleotide pools, which could impact chromatin states at specific phases of the cell cycle. Such approaches could induce metabolic alterations and/or DNA damage, which could reshape protein recruitment and histone modifications. In this protocol, we describe ways to fix and sort cells across the cell cycle based on their DNA content. We further detail immunoprecipitation and library preparation, allowing analysis of the epigenome by chromatin immunoprecipitation sequencing (ChIP-seq) for small numbers of cells. For complete details on the use and execution of this protocol, please refer to Van Rechem et al. (2021).

Original languageEnglish
Article number101243
JournalSTAR Protocols
Volume3
Issue number2
DOIs
StatePublished - Jun 17 2022

Keywords

  • Cell isolation
  • ChIPseq
  • Chromatin immunoprecipitation (ChIP)
  • Flow Cytometry/Mass Cytometry
  • Molecular Biology

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