Odd problem with RAPD
biochem2 at macqm.miyazaki-med.ac.jp
Wed Nov 2 05:58:39 EST 1994
> Robert Rumpf wrote:
> > I'm posting this for a friend...he's successfully amplified a RAPD band
> > and cloned it, then sequenced it using vector primers. He went ahead and
> > designed primers for the RAPD based on the sequences he obtained and now
> > can't amplify using these specific primers! Anyone have a problem similar
> > to this or perhaps have an explanation for it?
> I assume your friend used the clone as a positive control template and
> recovered the expected size band. Suggest to him to dilute the clone into
> something like salmon sperm DNA to simulate the amount and complexity of
> unique DNA in the target genome and see if that works. This will distinguish
> between a technical flaw in the amp. vs. the target seq. just not being there.
> Hope this helps.
> Steve Hardies, Dept. of Biochemistry, Univ. of Texas HSC at San Antonio
> Hardies at thorin.uthscsa.edu
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