GC advantage!

john f hess (remove-this) jfhess at ucdavis.edu
Mon Apr 21 11:56:48 EST 1997


I agree with the other reply.  We had tried everything to amplify a
region of our gene.  Different Taqs, different optimization techniques,
different co-solvents, everything!

Clontech GC advantage HAS NOT worked for every reaction we have tried it
on, BUT IT HAS BEEN THE BEST at the reactions we previously had no
success with.  Qiagen also makes a PCR kit that comes with "Q-solution"
and that worked OK for us.  However, there is one reaction that the
Clontech kit works with and the Qiagen kit doesn't.  At present, we are
running through reactions with the clontech kit to see what works and
cloning fragments.

BTW, the identity of the Qiagen Q solution is not revealed but I think it
is TMAC.  Ditto for the GC amp solution in the clontech kit.  However, I
know from previous work that in our attempts to get TMAC and Gibco Taq to
amplify our gene, we had no success.  We did have success with some
reactions with 5% formamide and lower annealing temps.  Also, we have 1
reaction of ~200 bp that only worked with formamide UNTIL we changed the
denaturation T to 98.  AT 98 denature (30 Secs), the reaction works well.

hope you find this interesting, 

cheers,


John F. Hess, PhD                     Phone me 916 752 8420
Dept of Cell Bio. and Hum. Anat.      FAX me 916-752-8520 (ask first)
University of California at Davis
Davis, CA 95616                       Email me jfhess at ucdavis.edu



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