need help with cloning by pcr

Margon Vandongen vando005 at mc.duke.edu
Tue Aug 29 15:04:19 EST 1995


Lisa,

Did you actually design the primers with oligo? I seem to 
remember that in the article about the program it warned
you against GC clamps.
I assume you know wether Vent works in taq buffer because
it does come with its own buffer that is different from the 
taq buffer.
Try going way down with your template concentration you are
using too much, about a ng or less should be fine.
I also assume you did buy your primers deprotected since
that would be a problem too.
Did you use either one of the primers with a primer that
you know works as a positive control?
You could easily see if the primers are uniformly made
by running an acrylamide gel (see red book)
Good luck,

Margon
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