Materials required for Cell cycle analysis by flow cytometer
- Ethanol 70%
- 100 micro litre Propidium iodide from solution 50 µg/ml)
- 100 micro litre Ribonuclease I (stock 100 µg/ml)
Steps
1
Harvest the cells in the appropriate manner and wash in PBS.
2
STEP 2 Fix in cold 70% ethanol.
3
Fix for 30 min at 4°C.
4
Wash 2 X in PBS.
5
Treat the cells with 50 µl of a 100 µg/ml stock of ribonuclease (RNase). This will ensure only DNA stainining.
6
Add 100 µl PI from 50 µg/ml stock solution
Wait for 2 to 5 min and take a reading.
