Improved flow cytometric analysis of the budding yeast cell cycle.
The budding yeast, Saccharomyces cerevisiae has been a remarkably useful model system for the study of eukaryotic cell cycle regulation. Flow cytometric analysis of DNA content in budding yeast has become a standard tool for the analysis of cell cycle progression. However, popular protocols utilizing the DNA binding dye, propidium iodide, suffer from a number of drawbacks that confound accurate analysis by flow cytometry. Here we show the utility of the DNA binding dye, SYTOX Green, in the cell cycle analysis of yeast. Samples analyzed using SYTOX Green exhibited better coefficients of variation, improved linearity between DNA content and fluorescence, and decreased peak drift associated with changes in dye concentration, growth conditions or cell size.
Haase
Steven B
SB
Department of Molecular Biology, MB7; The Scripps Research Institute; La Jolla, California 92037, USA. haase@scripps.edu.
Reed
Steven I
SI
D002453:Cell Cycle; D002455:Cell Division; D004396:Coloring Agents / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011419:Propidium / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology*
D004396:Coloring Agents; D005456:Fluorescent Dyes; D009930:Organic Chemicals; C402795:SYTOX Green; D011419:Propidium