PI staining Cell Cycle Progression[DNA staining]

Fix Cells

1. Trypsinize cells (if adherent) and transfer to microcentrifuge tube
2. Centrifuge at 10,000 rpm for 10 sec. and aspirate supernatant
3. Flick tube to loosen pellet (important to avoid clumps)
4. Add 500 uL 70% EtOH dropwise, while vortexing
5. Store samples overnight at -20'C

Stain Cells

1. Spin cells and aspirate supernatant as above
2. Flick tube to loosen pellet and add 500 uL PBS to rinse cells
3. Spin cells ans aspirate supernatant and resuspend in 400 uL PI solution (1/10 volume 1 mg/mL propidium iodide, 1/10 volume 10 mg/mL RNase A, 8/10 volume PBS) and transfer sample to flow tube
4. Incubate 30 min. at 37'C in the dark
5. Run samples on FACSCAN