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Updated: Jun 4, 2026
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
Analyzing DNA Replication I: Labeling Animals, Tissues, and Cells with Bromodeoxyuridine (BrdU)
CSH Protocols
|March 2, 2011
Summary
Detecting cells in S-phase of the cell cycle is crucial for understanding cell growth and cancer. This study details a method using 5-bromodeoxyuridine (BrdU) incorporation for precise S-phase identification in various biological samples.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Cell cycle progression and S-phase detection are vital for assessing cell growth and tumorigenicity.
- Existing markers like DNA polymerase and proliferating cell nuclear antigen lack S-phase specificity.
- 5-bromodeoxyuridine (BrdU) incorporation offers a more precise method for S-phase identification.
Purpose of the Study:
- To provide a detailed protocol for detecting S-phase cells using BrdU incorporation.
- To enable precise identification of DNA replication sites in various biological contexts.
Main Methods:
- Incorporation of modified DNA precursors, specifically 5-bromodeoxyuridine (BrdU), into cellular DNA.
- Detection of incorporated BrdU using fluorochrome- or enzyme-coupled antibodies.
- Application of the method to cultured cells, isolated tissues (in vitro), and whole animals (in vivo).
Main Results:
- BrdU is phosphorylated and incorporated into DNA, replacing deoxythymidine triphosphate.
- Labeled cells exhibit discrete nuclear sites (foci) corresponding to DNA replication sites.
- Distinct patterns of DNA replication foci characterize different stages of S-phase.
Conclusions:
- BrdU labeling provides a rapid and precise method for identifying S-phase cells.
- The protocol is adaptable for in vivo and in vitro studies across different biological sample types.
- This technique enhances the study of cell proliferation and DNA replication dynamics.
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