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Yeast Cell Synchronization by Nocodazole

Nocodazole inhibits microtubule dynamics thus disrupting mitotic spindle function. Blocks the cell cycle at G2/M phase. Therefore, cells will arrest with large buds and have 2N DNA content.

  1. Grow yeast cells in YPD. Add dimethyl sulfoxide (DMSO) to a final concentration of 1% just before adding Nocodazole at the final concentration of 15 microgram/ml.
    Adding DMSO helps nocodazole to more efficiently arrest cells, because Nocodazole dissolves better in DMSO.
  2. Incubate yeast at 30 C for 1.5 to 3 h.
  3. 90 min after the addition of Nocodazole, start checking yeast cells under the microscope. Examine every 30 min afterwards. Count the number of arrested cells. It should be more than 95%. Arrested cells will have buds almost as big as mother cells.
    Note: Sensitivity of every strain to Nocodazole may be different. It needs to be tested. Too little or too much Nocodazole will not give well synchonized populations.
  4. Good luck!

Scholarly Articles on Yeast Cell Synchronization

Day A, Schneider C, Schneider BL. Yeast cell synchronization. Methods Mol Biol. 2004;241:55-76.