degenerate PCR

Emma Macfarlane emma at HERA.MED.UTORONTO.CA
Mon Jun 27 10:48:14 EST 1994


Phil, I recently did a similar amplification using six codon primers with
2 - 4 fold degeneracy but I included inosine where degeneracy was 4 fold
The primers consequently had very low Tm's and the annealing step in the 
PCR had to be kept at 37oC but after 35 cycles I had enough product to 
gel purify and subclone.  I did however get alot of non-specific products
but since I knew the size of the correct product and could gel-purify it
easily this wasn't a problem.
Good luck
Emma



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