


Cell Cycle Analysis Protocol - PI Staining
- Harvest cells
washing in PBS.
- Fix pelleted cells
in ice-cold 70% ethanol by adding with a pasteur pipette on a vortex.
Leave cells at 4oC
from 30 mins to a week.
- Pellet cells at
approximately 2,000 rpm for 5 mins.
- Wash
twice in PBS.
- Add 50 ml RNAse to pelleted cells (100 mg/ml
Sigma R5125) and incubate at RT or 37oC for 15 mins.
- Add 200-400 ml of PI (final concentration 50 mg/ml
Sigma P4170).
- Analyse by flow cytometry
collecting 25,000 events per sample.