32P decay and kinase reactions

Bruce Byrne byrne at soma.umdnj.edu
Fri Jul 31 15:49:18 EST 1992


Tom Schneider wrote, regarding the 32P decay issue, that most
molecular biologists don't care what the decay products are.  Maybe
so, but we should.  Who, following these threads, can solve my
puzzle for me.

1.  On Friday I receive my high specific activity (6000 Ci/mmol)
gamma 32P from NEN.  I use a kinase reaction to end label oligo and
probe some blots (OK, maybe on Monday).  Hot stuff.  Good specific
activity.
2.  Two weeks later, (or three) I again need to probe.  By now my
oligo is half (or less) the specific activity, as is my gamma. 


Questions not resolved by calls to NEN or Promega or even very
limited trials at the bench.

1.   Is the ATP + ADP/S theoretical soup that I have in my NEN
container a good substrate for kinase?  Yes, I know it works, use
it all the time at that age.  But can the kinase distinguish
between its destiny (making great probe) and the ADP/S stuff?

2.   Do the chemical byproducts of decay inhibit a kinase reaction?

Trials at the bench have limited success in my hands since I have
not found the kinase reaction to be particularly quantitative.  Any
thoughts?

Bruce
Please respond directly to: byrne at umdnj.edu

The University of Medicine and Dentistry of New Jersey (USA)



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