33P: results!

jmv at canctr.mc.duke.edu jmv at canctr.mc.duke.edu
Fri Oct 9 14:16:23 EST 1992

Here, finally, is what I've found out about 33P. It works terrifically
with regular (2 primer) "hot" PCR, using the same parameters as with
32P. Now we must try to titer reactions to see whether the 33P can be
diluted by a factor large enough to offset the price vs. 32P.

For cycle sequencing (gamma 33P end-labelling the primer), 33P used
in the same amount & same conditions as 32P yields much sharper results,
though so light that exposure time is at least doubled. The amount ued
according to Promega fmol (tm) kit methods is so small that, since 33P
1/2-life is ~4x as long as 32P's, we get a better deal paying 2x as much.

Q: Has anyone used 33P for probe labelling (for Southerns, eg)?
Q2: Has anyone had success with 33P and the CircumVent cycle sequencing
kit? There's a NEB promo for the latter offering free 33P, & I'm wondering
if it's worth anything.


DB Loeb
-Neurology Div., Duke U. Med. Ctr.
--Durham, NC  27710

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