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

STAR Protoc. 2022 Mar 15;3(2):101243. doi: 10.1016/j.xpro.2022.101243. eCollection 2022 Jun 17.

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).

Keywords: Cell isolation; ChIPseq; Chromatin immunoprecipitation (ChIP); Flow Cytometry/Mass Cytometry; Molecular Biology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle / genetics
  • Chromatin Immunoprecipitation / methods
  • Chromatin Immunoprecipitation Sequencing*
  • Chromatin* / genetics
  • Histone Code

Substances

  • Chromatin