Looking for a PCR P32-labelling protocol

Amy Anne Caudy aacaudy at artsci.wustl.edu
Mon Aug 3 08:55:43 EST 1998


Well, I just do a "normal" PCR reaction but use normal concentrations of 3
nucleotides and 1/10th concentration of the 4th, plus 5-7 ul of the
alpha-labelled 4th nucleotide. Choose your conditions normally for your
primers.

Such probes have worked fine for me in Southerns. 

- Amy Caudy
Washington University 

m.champagne at cellbio.duke.edu wrote:
: Hi,

: I'm stuck with a piece of DNA that won't label well when I use random-primed
: labelling.  I tried Prime-it kit with P32, B-M's Digoxygenin's kit, homemade 
: Klenow version of random-primed labelling...  My control probes incorporate
: the label pretty well, but my piece of DNA won't.

: I've tried a PCR-based P32 incorporation once, but it didn't work, and before
: i try again, I thought I'd ask for general advice on labelling, and
: also for comments from people who've used PCR-based P32 labelling.

: Thanks,

: m.champagne at cellbio.duke.edu



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