MFox at vines.ColoState.EDU
Mon Mar 4 12:09:52 EST 1996
Dr. Darley wrote concerning different methods for BrdUrd processing and
wondered whether comparisons have been made between methods. Dolbeare and
Gray, Cytometry 9, 631-635, 1988, used a method of Eco R1 endonuclease plus
Exo III exonuclease to open up single stranded regions of the DNA and found
it to be more sensitive than HCl denaturation. I have also used both methods
extensively and found that a much shorter pulse of BrdUrd could be used with
the Eco/Exo method than with HCl. I was using image analysis to look at
BrdUrd incorporation and the results were better with the Eco/Exo method.
However, for flow I routinely use the HCl method because it is cheaper and
easier and works fine for what we normally do.
Colorado State University
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