DMSO vs. raising annealing temp. in PCR
mmorales at acpub.duke.edu
Fri Dec 20 12:38:17 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 seems to be rather widely used to increase the specificity of PCR,
: but I have always thought it may be replaced with tricks in annealing/
: denaturation temperature and reagent concentrations.
In my experience, PCR conditions a very empirical. Each template and primer
pair can present unique problems. So I guess what I am saying is that it is
a thing to try, and it will either help or it won't. There are a lot of
helpful articles on PCR optimization, E-mail me and I'll send you a list, if
: Am I wrong?
: May DMSO be better in increasing the specificity of PCR (and the yield of
: the right product) than the other tricks?
: Is anyone with experiences or knowledge willing to share them?
: Timo Hiltunen
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