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Related Experiment Video

Updated: May 19, 2026

Assessment of Global DNA Double-Strand End Resection using BrdU-DNA Labeling coupled with Cell Cycle Discrimination Imaging
06:44

Assessment of Global DNA Double-Strand End Resection using BrdU-DNA Labeling coupled with Cell Cycle Discrimination Imaging

Published on: April 28, 2021

A high-throughput, flow cytometry-based method to quantify DNA-end resection in mammalian cells.

Josep V Forment1, Rachael V Walker, Stephen P Jackson

  • 1The Gurdon Institute and Department of Biochemistry, University of Cambridge, CB2 1QN Cambridge, United Kingdom. j.forment@gurdon.cam.ac.uk

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|August 16, 2012
PubMed
Summary

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We developed a high-throughput flow cytometry method to quantify Replication Protein A (RPA) recruitment. This technique enables unbiased measurement of cell proliferation and DNA repair by homologous recombination (HR).

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Replication protein A (RPA) is crucial for DNA metabolism, binding single-stranded DNA (ssDNA) during replication and repair.
  • RPA localization to DNA damage sites indicates DNA-end resection and homologous recombination (HR) activity.
  • Current methods for quantifying RPA recruitment are labor-intensive and analyze small cell populations.

Purpose of the Study:

  • To develop a high-throughput, quantitative method for measuring RPA recruitment.
  • To establish flow cytometry as a tool for assessing cell proliferation and DNA repair via HR.

Main Methods:

  • Utilized RPA staining in conjunction with flow cytometry.
  • Developed a method for high-throughput, quantitative analysis of RPA recruitment.

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Visualizing Single-Stranded DNA Foci in the G1 Phase of the Cell Cycle
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Visualizing Single-Stranded DNA Foci in the G1 Phase of the Cell Cycle

Published on: December 22, 2023

Analysis of DNA Double-strand Break (DSB) Repair in Mammalian Cells
13:10

Analysis of DNA Double-strand Break (DSB) Repair in Mammalian Cells

Published on: September 8, 2010

Related Experiment Videos

Last Updated: May 19, 2026

Assessment of Global DNA Double-Strand End Resection using BrdU-DNA Labeling coupled with Cell Cycle Discrimination Imaging
06:44

Assessment of Global DNA Double-Strand End Resection using BrdU-DNA Labeling coupled with Cell Cycle Discrimination Imaging

Published on: April 28, 2021

Visualizing Single-Stranded DNA Foci in the G1 Phase of the Cell Cycle
08:30

Visualizing Single-Stranded DNA Foci in the G1 Phase of the Cell Cycle

Published on: December 22, 2023

Analysis of DNA Double-strand Break (DSB) Repair in Mammalian Cells
13:10

Analysis of DNA Double-strand Break (DSB) Repair in Mammalian Cells

Published on: September 8, 2010

Main Results:

  • The developed flow cytometry method allows for unbiased and quantitative measurement of RPA recruitment.
  • This technique can be used to assess both cell proliferation and DNA repair by HR.

Conclusions:

  • Flow cytometry offers a significant advancement over traditional microscopy for quantifying RPA recruitment.
  • This method provides a powerful tool for high-throughput analysis of DNA repair and cell proliferation.