P32 vs. P33 for PCR detection
Dima Klenchin
klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu
Mon Jul 2 19:55:56 EST 2001
mwcrepeau at hotmail.com (Marc Crepeau) wrote:
:Why do people sometimes use P33 to label PCR products for detection
:via autoradiograms? What is the advantage over P32? We order P32
:routinely for our Southern blot hybridizations, and I'm told it's
:cheaper than P33. But I have some microsatellite markers that I want
:to use, and the information I have states that the developers employed
:P33 for detection, not P32 (the species in question is highly
:polyploid, so visualization with EtBr is not sufficiently sensitive).
:Would I get the same results if I just spiked the PCR with the alpha
:labeled P32 that we already have in the lab?
P33 is "cleaner". Less energy = smaller effective angle of
radiation scatter (see pic) = sharper bands = better resolution,
which in many applications is more important than sheer
sensitivity.
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