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Determination of S-Phase Duration Using 5-Ethynyl-2′-deoxyuridine Incorporation in Saccharomyces cerevisiae
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Determination of S-Phase Duration Using 5-Ethynyl-2′-deoxyuridine Incorporation in Saccharomyces cerevisiae

Determination of S-Phase Duration Using 5-Ethynyl-2′-deoxyuridine Incorporation in Saccharomyces cerevisiae

DOI:
10.3791/64490-v

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08:40 min

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October 21, 2022

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, ,
1Institut de Génétique Moléculaire de Montpellier,Université de Montpellier, CNRS

Chapters

  • 00:05Introduction
  • 00:42Saccharomyces cerevisiae Cell Culture
  • 02:00EdU Labeling
  • 02:40Cell Fixation and Permeabilization
  • 03:20Click-It Reaction for Cytometry Analysis
  • 06:32Results: Using EdU Labeling to Determine the S-Phase Duration in Saccharomyces cerevisiae
  • 07:56Conclusion

Summary

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We describe two complementary protocols to accurately determine S-phase duration in S. cerevisiae using EdU, a thymidine analog, which is incorporated in vivo and detected using Click chemistry by microscopy and flow cytometry. It allows for the easy characterization of the duration of DNA replication and overlooked replication defects in mutants.

Tags

Keywords: S-phase Duration5-Ethynyl-2′-deoxyuridineSaccharomyces CerevisiaeCell Cycle RegulationSynchronized Budding YeastAlpha FactorEDU LabelingPI-EDU Bivariate Plot

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