UNIT 11.13 Cell Cycle Analysis of Yeasts
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Cytometry
How to Cite
Fortuna, M., João Sousa, M., Côrte-Real, M., Leão, C., Salvador, A. and Sansonetty, F. 2001. Cell Cycle Analysis of Yeasts. Current Protocols in Cytometry. 13:11.13:11.13.1–11.13.9.
- Published Online: 1 MAY 2001
- Published Print: JUL 2000
Staining protocols generally designed for the flow cytometric analysis of the cell cycle in mammalian cells are frequently not satisfactory for quantification of the various cell-cycle phases in yeasts. High CVs limit the accuracy of DNA content measurement and estimates of populations in cell-cycle compartments. This unit describes a staining procedure for yeasts using the sensitive nucleic acid stain SYBR Green I, which binds to double-stranded DNA with high selectivity and which has a much higher fluorescence quantum yield upon binding than most other commonly used fluorophores. The properties of this dye combined with optimized sample processing provide high-resolution DNA analysis, with half-peak CVs around 3 to 4% and clear-cut identification of the S phase.