Cell Cycle Analysis Protocol - 7-AAD Staining

1. Harvest cells washing in PBS.
2. Fix pelleted cells in ice-cold 70% ethanol by adding with a pasteur pipette on a vortex. Leave cells at 4^oC from 30 mins to a week.
3. Pellet cells at approximately 2,000 rpm for 5 mins.
4. Wash twice in PBS.
5. Add 200-400 ml of 7-AAD (final concentration 25 mg/ml Sigma).
6. Analyse by flow cytometry collecting 25,000 events per sample.