hygromycin primers

WS via methods%40net.bio.net (by novalidaddress from nurfuerspam.de)
Tue Oct 9 17:16:34 EST 2007


Dear SVemur... from gmail.com

Basically, I would not give too much on calculated annealing
temperatures. I'd simply check the reaction in a gradient cycler (if
possible, else just run a set of samples prepared from one Rxn mix
with different Ta) where you may get an idea about at which annealing
temperatures your product looks best. If you have that HygR gene /
cDNA somewhere in a plasmid, too, you might use that as well for your
studies (in parallel) at a sub-maximal dilution (if not yet, simply
clone your PCR product into e.g. a TA vector). Will be a nice positive
control for the PCR system itself, including the primers.

Best regards. Wo



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