Protocol for cell cycle analysis by flow cytometer

 

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.