Odd problem with RAPD

Tue Nov 1 15:30:09 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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