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
Such probes have worked fine for me in Southerns.
- Amy Caudy
m.champagne at cellbio.duke.edu wrote:
: 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.
:m.champagne at cellbio.duke.edu