È necessario avere un abbonamento a JoVE per visualizzare questo. Accedi o inizia la tua prova gratuita.
Capitoli
- 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
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.
