PCR labeled probes

Kerry Fulcher kfulcher at ptloma.edu
Fri Jul 23 11:10:06 EST 1999


Does anyone out there generate radiolabeled probes via PCR.  I 
have done
this in the past to make probes for Northerns etc... but now seem 
to be
having troubles.  I was wondering if anyone has a protocol for doing 
this
that I can compare with so see where my problems could be 
coming from.

In the past I have simply added the 3 cold dNTPs in at typical 
levels and
added 5 ul of P32 dCTP and cycled away.  This used to generate 
nice probes
with high specific activity.... but lately I am not getting very good
incorportation (even though the non-radioactive controls have nice
yields).
 Any suggestions?

Thanks,

Kerry
-^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Kerry Fulcher
Biology Department, Point Loma Nazarene University
San Diego CA,  92106
Phone = 619-849-2651                         
Fax =   619-849-2598                  
E-mail = kfulcher at ptloma.edu

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"Everything can be taken from a man but one thing: the last of the human
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