Problems with compDNA

SerAln SerREMOVETHISAln at netscape.net
Wed Mar 13 05:06:49 EST 2002


And what about using just one primer ?
I never tried this, but sometimes, when one of the primers in a normal PCR is 
not working we get a lot of what we consider ssDNA.

Sergio

Tom Knight wrote:

> "knud" <kknald at hotmail.com> writes:
> 
>>Do anybody know how to get rid of the complementary DNA strand after
>>PCR. My upper strand is Biotinylated, and I would be really happy to
>>get rid of the non biotinylated complementary strand since it is
>>causing me trouble in my DNA:DNA hybridization. Is there somekind of
>>Matrix I can use??? Thanks in advance.  Mads Hollegaard
>>
> 
> You might consider asymmetric PCR, with a large excess of one of the
> two primers.  In your case, the biotinylated primer could be in
> excess.  When the low concentration primer is exhaused, each cycle
> then performs a linear amplification of the template, yielding excess
> of the biotinylated version.  This won't get rid of all of the
> complementary strand, but might be good enough to solve your problem.
> 
> Of course, you could also bind the biotin, denature, and wash the
> unbound DNA, but it is problematic to then unbind it.
> 
> 




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