| Autor: |
Rosebrock AP; Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario M5S 3E1, Canada. |
| Jazyk: |
angličtina |
| Zdroj: |
Cold Spring Harbor protocols [Cold Spring Harb Protoc] 2017 Jan 03; Vol. 2017 (1). Date of Electronic Publication: 2017 Jan 03. |
| DOI: |
10.1101/pdb.prot088740 |
| Abstrakt: |
DNA synthesis is one of the landmark events in the cell cycle: G 1 cells have one copy of the genome, S phase cells are actively engaged in DNA synthesis, and G 2 cells have twice as much nuclear DNA as G 1 cells. 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.
(© 2017 Cold Spring Harbor Laboratory Press.) |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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