Synchronization of cell populations in G1/S and G2/M phases of the cell cycle.
The method described in the following chapter utilizes a double thymidine block (an inhibitor of DNA synthesis) followed by treatment of cells with nocodazole (a mitotic inhibitor) to obtain large cell populations at distinct phases of the cell cycle. Treatment with double thymidine results in a G1/S-phase arrested cell population, and the use of flow cytometry allows progression of the cells through the cell cycle to be monitored. Flow cytometry enables the calculation of timings for collection of cells at distinct cell cycle phases from G1/S (following treatment with thymidine) through to G2/M (owing to the presence of nocodazole).
Harper
Jane V
JV
School of Animal and Microbial Sciences, University of Reading, Reading, UK.
D000818:Animals; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000378:metabolism; D002460:Cell Line; D003584:Cytological Techniques / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015739:Nocodazole / Q000494:pharmacology; D051381:Rats; D013936:Thymidine / Q000378:metabolism / Q000494:pharmacology
D018797:Cell Cycle Proteins; D015739:Nocodazole; D013936:Thymidine