title: Cell cycle analysis of yeasts.
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.
authors.affiliation: Universidade do Minho, Braga, Portugal.
authors.forename: M
authors.initials: M
authors.lastname: Fortuna
authors.forename: M J
authors.initials: MJ
authors.lastname: Sousa
authors.forename: M
authors.initials: M
authors.lastname: Côrte-Real
authors.forename: C
authors.initials: C
authors.lastname: Leão
authors.forename: A
authors.initials: A
authors.lastname: Salvador
authors.forename: F
authors.initials: F
authors.lastname: Sansonetty
mesh_terms: D002453:Cell Cycle; D002469:Cell Separation / Q000379:methods*; D004247:DNA / Q000032:analysis / Q000737:chemistry; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000494:pharmacology; D015203:Reproducibility of Results; D015003:Yeasts / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D004247:DNA