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