Measuring DNA Content by Flow Cytometry in Fission Yeast

  • Sarah A. Sabatinos
  • Susan L. Forsburg
Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 521)

Summary

Flow cytometry is an essential tool to monitor DNA content and determine cell cycle distribution. Its utility reflects the relative ease of sample preparation and the stochiometric nature of the most popular DNA-binding dyes (propidium iodide and Sytox Green). Mammalian precedents using flow cytometry for replication and cell biology studies are attractive examples for S. pombe researchers. However, the study of DNA replication with multicolor analysis has lagged behind that in mammalian cells. We present basic and advanced protocols for analysis of DNA replication in fission yeast by flow cytometry including whole cell, nuclear “ghosts,” and two-color imaging with BrdU.

Key words

DNA replication Flow cytometry (FACS) DNA content DNA replication BrdU detection Immunofluorescence 
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Notes

Acknowledgments

Thanks to David Chambers of the Center for Cell and Molecular Imaging at the Salk Institute for much helpful advice, and Oscar Aparicio at USC for sharing his FACScan. Supported by grant NIH R01 GM59321 to SLF.

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Sarah A. Sabatinos
    • 1
  • Susan L. Forsburg
    • 1
  1. 1.Department of Molecular and Computational BiologyUniversity of Southern CaliforniaLos AngelesUSA

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