Analyzing Schizosaccharomyces pombe DNA Content by Flow Cytometry
| Autor: | Trond Stokke, Christiane Rothe, Silje Anda, Erik Boye, Beáta Grallert |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
medicine.diagnostic_test biology Chemistry Cell cycle Flow Cytometry biology.organism_classification Fluorescence General Biochemistry Genetics and Molecular Biology Yeast Flow cytometry Cell biology 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology Schizosaccharomyces Schizosaccharomyces pombe Content (measure theory) medicine DNA Fungal DNA Cytokinesis |
| Zdroj: | Cold Spring Harbor Protocols. 2016:pdb.prot091280 |
| ISSN: | 1559-6095 1940-3402 |
| DOI: | 10.1101/pdb.prot091280 |
| Popis: | Flow cytometry can be used to measure the DNA content of individual cells. The data are usually presented as DNA histograms that can be used to examine the cells’ progression through the cell cycle. Under standard growth conditions, fission yeast cells do not complete cytokinesis until after G1 phase; therefore, DNA histograms show one major peak representing cells in G1 (2×1C DNA) and G2 phase (1×2C DNA). By analysis of the duration of the fluorescence signal as well as the intensity of the DNA-related signal, it is possible to discriminate between cells in M/G1, S, and G2. This protocol describes how to prepare cells for flow cytometry and analyze them. We also describe the application of barcoding for more accurate comparison of samples. |
| Databáze: | OpenAIRE |
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