Double Thymidine Synchronizing Protocol

Double Thymidine Synchronizing Protocol:

Preparation:
1. Complete Culture Medium
2. 100mM Thymidine in complete medium
3. Phosphate Buffer Saline ( PBS )

Protocol:
1. Seeding cell on 40mm culture dish. Incubate at 37 degree until 40% confluency.
2. Add 40ul 100mM thymidine solution. Final concentration is 2mM.
3. Incubate 24Hrs.
4. Wash with PBS. Add complete medium without thymidine.
5. Incubate 8Hrs.
6. Add 40ul 100mM thymidine solution. Final concentration is 2mM.
7. Incubate 24Hrs.
8. Wash with PBS. Add complete medium without thymidine.
9. Ready for experiment.