DMSO vs. raising annealing temp. in PCR
jkropp at itsa.ucsf.edu
Mon Dec 23 18:26:22 EST 1996
> : I am trying to amplify a fragment of about 100 bp but
> : am getting a smear of products with only little of the desired one.
> : I am not using too many cycles and have raised the annealing
> : temperature to its highest limit.
DMSO may help at 5-10%. Or other optimization tricks, like titrating Mg.
If that doesn't cut it, grit your teeth design a new primer pair.
I've found the "internal stability" parameter in the Oligo primer analysis
application to be very useful for avoiding promiscuous primers.
"no brains...no headache!"
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