Hot PCR

thobman at anat.med.ualberta.ca thobman at anat.med.ualberta.ca
Fri Jun 21 04:56:48 EST 1996


Does anyone have a protocol for producing a 32-P-labeled PCR product 
(approx. 100 bp) directly from a library.  Instead of cloning the PCR 
product an then labeling it for use as a probe (for a Northern blot), 
we would like to directly amplify the fragment using alpha-32-P-dATP 
to generate a hot probe.  We need to know what ratio of hot dATP/cold 
dATP to use in the reactions and suggested # of cycles and whether or 
not it is necessary to purify the product before use (other than 
Nunc-trap columns)

Thanks

Tom Hobman



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