Re: Staining with PI & BrdU
kukuruga%kasle1.dnet.wayne.edu@rocdec.roc.wayne.edu
Sat, 30 Mar 1996 10:52:13 -0500
Someone recommended shortening your pulse time. . . I agree. 2 hours in
your cells is probably too long. 30 minutes should be sufficient. As far
as %s-phase is concerned, however, it's not a simple matter of doing the
incorporation and looking for the numbers. Several references are available
for your guidance -- look them up. Two points; 1) straight cell cycle
estimation by PI-DNA histogram analysis should provide a reasonable
comparison for your BrdU data, 2) perhaps as important as a technical issue
is the point in the growth curve aat which you pulse and assay your cell culture
and assay your cell culture. PI-DNA analysis of cell cultures over a period
of days (or even hours) can help you determine appropriate analysis time-
points.
MAK.
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