Reinhard Kunze (kunze at gen1.genetik.uni-koeln.de) wrote:
> Is there everyone having experience in the use of degenerated oligonucleotides?
> My plan is to order a 60 bases long oligonucleotide for site-directed
> mutagenesis, which should have four wobble-positions. Could you please send
> me your advice, in which ratios
First, think about having your mutations near the center of your oligo. By the
size of your oligo, I figure you'll be using the Kunkel method. I have more
experience with PCR mutagenesis.
Obviously, the more degenerate your oligos, the greater number of different
oligos you'll be using, and the lower the concentration of each population of
oligo. If you can avoid degeneracy and can pre-select your intended mutations,
the easier this experiment will be.
Benoit_Hebert at iaf.uquebec.ca
Virology Research Center
Laval, Quebec, Canada H7N 4Z3
phone: (514) 687-5010, ext. 4636
fax: (514) 686-5626