FACS
DA Wolf, September 06
- Trypsinize cells from 10 cm or 15 cm dish and wash in
media to inactivated trypsin
- Wash
in 5 ml PBS
- Resupsend in 600 ul PBS
- Add
1.4 ml 100% cold ETOH (-20C) while vortexing
- Fix
overnight at -20C
- Wash
in 5 ml PBS
- Resuspend in 500 ul PBS, 2.5 ug RNAse A (from 10 mg/ml
stock, preboiled)
- Incubate
at RT for 45 min
- Add 40
ug/ml propidium iodide (from 1 mg/ml stock)
- Run on
FACS