title: Analysis of the Budding Yeast Cell Cycle by Flow Cytometry.
abstract: DNA synthesis is one of the landmark events in the cell cycle: G1cells have one copy of the genome, S phase cells are actively engaged in DNA synthesis, and G2cells have twice as much nuclear DNA as G1cells. Cellular DNA content can be measured by staining with a fluorescent dye followed by a flow-cytometric readout. This method provides a quantitative measurement of cell cycle position on a cell-by-cell basis at high speed. Using flow cytometry, tens of thousands of single-cell measurements can be generated in a few seconds. This protocol details staining of cells of the budding yeast Saccharomyces cerevisiae for flow cytometry using Sytox Green dye in a method that can be scaled widely-from one sample to many thousands and operating on inputs ranging from 1 million to more than 100 million cells. Flow cytometry is preferred over light microscopy or Coulter analyses for the analysis of the cell cycle as DNA content and cell cycle position are being directly measured.
authors.affiliation: Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario M5S 3E1, Canada.
authors.forename: Adam P
authors.initials: AP
authors.lastname: Rosebrock
mesh_terms: D002453:Cell Cycle; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000502:physiology*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes