DNA labeling alternatives

Klaus Salger salger at wap4.zi.biologie.uni-muenchen.de
Tue May 23 23:23:29 EST 1995


marivonne (marivonne at bio.tamu.edu) wrote:
: Hello all,

: I am interested in using a PCR fragment (approx. 600 bp) as a probe in
: Southern blots. In order to label it (32P), which procedure would be more
: efficient, nick translation or random-primed labeling? What are the pro's
: and con's of nick translating a PCR fragment for use as a probe? Pro's and
: con's of random-primed labeling for same purpose?

: Any advice will be *greatly* appreciated. Thank you.

: Marivonne Rodriguez
: (marivonne at bio.tamu.edu)

I have allways used random priming with good results. It's simple, fast
and gives good labeling efficiency.
But in your case I would use the PCR primers rather than random primers
for labeling. If you know which one is the coding strand you could even
use just one of them to end up with a strand specific probe.
I think this should work very good but I've never tried it (didn't have
the primers) so I couldn't compare random vs. "specific" priming.

Cheers
  Klaus
 
--
Klaus Salger                phone : ++49 (0)89 5902 -502
Zoologisches Institut       FAX   :                 -450
AG MacWilliams              e-mail: salger at zi.biologie.uni-muenchen.de
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