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

Abstract

Summary: 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).

Keywords

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