From: Aislinn D. Black (aislinn.black@alumni.carleton.edu)
Date: Thu Jun 19 2003 - 16:02:02 EST
Hello flow-ers, My colleague and I have a question about some cell cycle analysis in rat myocytes. We're using a fairly scaled-back protocol from what I've used in other cell types, as we were losing too many cells in the prep otherwise, but we just wanted to see if others in the list had used something similar, and had any feedback. After treatment, cells are harvested from their dishes with trypsin, neutralized, and centrifuged. The pellet is then resuspended in PBS and 70% ethanol, and frozen at -20C. After being frozen for varying lengths of time (from two days to a week), they are then centrifuged and resuspended in a staining buffer comprising .1% Triton X, 50ug/mL PI, and 50 ug/mL RNase A in PBS. (From Communal C et al., 1998. Circulation 98:1329-1334). Thanks, Aislinn Aislinn D. Black NY Harbor VA Medical Center - Brooklyn Campus black.aislinn@ny-harbor.med.va.gov 718-836-6600 x3826 -- http://www.fastmail.fm - Consolidate POP email and Hotmail in one place
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