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

Jianming Wang*, Marco Saponaro

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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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
Issue number2
Early online date29 May 2021
Publication statusPublished - 18 Jun 2021

Bibliographical note

Funding Information:
We thank the Genomics Birmingham for help with NGS library preparation and sequencing samples and Paola Pietroni and Adriana Flores-Langarica from the Flow Cytometry Facility for sorting mitotic cells, all from the University of Birmingham. This work was supported by a University of Birmingham fellowship, the Wellcome Trust ( 202115/Z/16/Z ), the Royal Society ( RG170246 ), and the BBSRC ( BB/S016155/1 ) to M.S.


  • 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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