Cell Cycle Analysis of Yeasts

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Abstract
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.