Measuring DNA content by flow cytometry in fission yeast.
Flow cytometry is an essential tool to monitor DNA content and determine cell cycle distribution. Its utility in fission yeast reflects the ease of sample preparation, the stochiometric binding of the most popular DNA dyes (propidium iodide and Sytox Green), and ability to monitor cell size. However, the study of DNA replication with multicolour flow analysis has lagged behind its use in mammalian cells. We present basic and advanced protocols for analysis of DNA replication in fission yeast by flow cytometry including whole cell, nuclear "ghosts," two-color imaging with BrdU, and estimates of DNA synthesis using EdU.
Department of Molecular and Computational Biology, University of Southern California, 1050 Childs Way, RRI 108, Los Angeles, CA, 90089-2910, USA.
Sarah A
SA
Sabatinos
Susan L
SL
Forsburg
D001973:Bromodeoxyuridine / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D004271:DNA, Fungal / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods*; D016196:S Phase; D012568:Schizosaccharomyces / Q000378:metabolism*; D013194:Staining and Labeling
D004271:DNA, Fungal; D001973:Bromodeoxyuridine