Nocodazole arrest cells in mitotic phase. Nocodazole arrest may be combined with with thymidine block by arresting cells after the first round of thimidine block.

Mammalian cells

1. Grow cells to sub-confluent.

2. Add nocodazole to a final concentration of 50 ng/ml.

3. Incubate for 14-18 hours.

Yeast

1. Grow cells to the mid-log phase.

2. Add nocodazole to a final concentration of 15-20 mg/ml. Nocodazole stock (15-20 mg/ml) is made with DMSO. 1% DMSO should be added to the control cells.

3. Return cells to the incubator. After 2 hours, 80% of nocodazole-treated cells should be large budded.

4. Nocodazole is not effective in arresting S. pombe cells.