Detecting Cell Cycle Stage and Progression in Fission Yeast, Schizosaccharomyces pombe. 2022

Zohreh Kianfard, and Kyle Cheung, and Daniel Rappaport, and Sirasie P Magalage, and Sarah A Sabatinos
Molecular Science Program, Yeates School of Graduate Studies, Toronto Metropolitan University (formerly Ryerson University), Toronto, ON, Canada.

We have previously described methods to synchronize cultures of fission yeast, Schizosaccharomyces pombe. In this chapter, we provide methods to detect cell cycle stage in cells and populations of S. pombe. These protocols used fixed samples. First, we describe sample preparation for flow cytometry of bulk DNA content. This technique allows users to monitor progression of DNA replication and detect any perturbation during the synthesis (S) phase of the cell cycle. Second, we describe methods to stain nuclei and septa of fixed yeast cells, and monitor proportions of cell cycle stages within cultures. Together, these methods provide the ability to compare cell cycle progression or delay between cultures, making use of the powerful molecular genetics tool that is S. pombe.

UI MeSH Term Description Entries
D002453 Cell Cycle The complex series of phenomena, occurring between the end of one CELL DIVISION and the end of the next, by which cellular material is duplicated and then divided between two daughter cells. The cell cycle includes INTERPHASE, which includes G0 PHASE; G1 PHASE; S PHASE; and G2 PHASE, and CELL DIVISION PHASE. Cell Division Cycle,Cell Cycles,Cell Division Cycles,Cycle, Cell,Cycle, Cell Division,Cycles, Cell,Cycles, Cell Division,Division Cycle, Cell,Division Cycles, Cell
D002455 Cell Division The fission of a CELL. It includes CYTOKINESIS, when the CYTOPLASM of a cell is divided, and CELL NUCLEUS DIVISION. M Phase,Cell Division Phase,Cell Divisions,Division Phase, Cell,Division, Cell,Divisions, Cell,M Phases,Phase, Cell Division,Phase, M,Phases, M
D004261 DNA Replication The process by which a DNA molecule is duplicated. Autonomous Replication,Replication, Autonomous,Autonomous Replications,DNA Replications,Replication, DNA,Replications, Autonomous,Replications, DNA
D012568 Schizosaccharomyces A genus of ascomycetous fungi of the family Schizosaccharomycetaceae, order Schizosaccharomycetales. Fission Yeast,Schizosaccharomyces malidevorans,Schizosaccharomyces pombe,Yeast, Fission,S pombe,Fission Yeasts
D016196 S Phase Phase of the CELL CYCLE following G1 and preceding G2 when the entire DNA content of the nucleus is replicated. It is achieved by bidirectional replication at multiple sites along each chromosome. S Period,Period, S,Periods, S,Phase, S,Phases, S,S Periods,S Phases

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