Staining with PI & BrdU

kukuruga%kasle1.dnet.wayne.edu@rocdec.roc.wayne.edu kukuruga%kasle1.dnet.wayne.edu at rocdec.roc.wayne.edu
Sat Mar 30 10:52:13 EST 1996


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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