Protocol for analysis of G2/M DNA synthesis in human cells

Jianming Wang, Marco Saponaro

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Abstract

G2/M DNA synthesis (G-MiDS) can be observed in one in five G2/M cells in unperturbed conditions by immunofluorescence microscopy. However, little is known of the genomic sites undergoing G-MiDS. Here, we describe a protocol which allows enriching for G2/M cells and investigating the sites of G-MiDS using BrdU-seq. This method can also be used to study the role of DNA replication or transcription-associated factors in affecting G-MiDS levels in different cell lines. For complete details on the use and execution of this protocol, please refer to Wang et al. (2021).

Original languageEnglish
Article number100570
JournalSTAR Protocols
Volume2
Issue number2
Early online date29 May 2021
DOIs
Publication statusPublished - 18 Jun 2021

Keywords

  • Cell Biology
  • Cell isolation
  • Flow Cytometry/Mass Cytometry
  • Molecular Biology
  • Sequencing

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)
  • Immunology and Microbiology(all)

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