Long PCR with degenerate primers and inosine
r.mistry at udcf.gla.ac.uk
Wed Nov 22 14:56:43 EST 1995
I would be very interested in hearing from anyone who has had
experience using the Gibco/BRL Elongase PCR kit (containing
pyrococcus thermostable polymerase and Taq). I have just tried it
out to amplify a ~12 kb product. However, I could not see any bands
at all, apart from primers. I'm using 30-mer degenerate oligos.
Also the primers contain inosine.
The PCR conditions were those recommended by the kit. Annealing
temperature of 55 C was used, which is known to give a range of
shorter non-specific products using Taq alone.
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