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Star Republic: Guide for Biologists

Synchronization of cells --- double thymidine block

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1. Grow cells to subconfluent (mid-log phase).

2. Add thymidine to a final concentration of 2 mM for 16-24 hours (one cell cycle).

3. Wash cells with PBS three times and re-feed with fresh grow medium.

4. Incubate for 10 hours before adding thymidine to a final concentration of 2 mM.

5. Incubate for 14-22 hours.

6. Repeat step 3. This will be time 0.