RAPD PCR breakdown

DrAnnie amyezers at kings.edu
Wed Mar 24 09:41:54 EST 2004


Thanks for some input.  

I just got the thermal cycler back from service where they replaced
the block -- didn't help.

We aren't using a hot start Taq (maybe we should?).  I ordered new Taq
to replace the quesitonable one.   But the Taq which was originally
Gibco is now sold by Invitrogen.  Can anybody suggest the best Taq for
RAPD's?

We have a 16S bacterial amplification that works beautifully on the
same machine but both our RAPD amplifications for beetles and fish are
not working.

We've just tried snap-freezing the primers, but I don't have those
results yet.  We do dilute those in water.

Thanks for the tip on the stored DNA -- haven't tried that yet.

Any other suggestions are welcome!



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