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Synchronizing yeast cultures
04/26/2012
I am struggling with synchronizing S. cerevisiae cells at G2/M with NocodazoleCan anyone offer some advice?
The BioTechniques Molecular Biology Forum brings scientists together in an online community to ask for and share advice regarding laboratory problems. This week’s highlighted question comes from the Cell Culture forum.
Q: I am struggling with synchronizing S. cerevisiae cells at G2/M with Nocodazole. I start with 5E6 cells and add Sigma nocodazole in DMSO at varying concentrations to titrate for the best concentration. I haven't found a single G2/M arrest with any of the concentrations I’ve tried. I incubate for at least 1 generation. Can anyone offer some advice?
A: For cell synchronization experiments, nocodazole is usually used at a concentration of 40-100 ng/mL of culture medium for 12-18 hrs. Prolonged arrest of cells in mitosis due to...
A: How long it takes for the cells to arrest depends on how quickly nocodazole depolymerizes…
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Keywords:
cell culture
