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Usage of degenerated oligonucleotides

Benoit_Hebert at IAF.UQUEBEC.CA Benoit_Hebert at IAF.UQUEBEC.CA
Mon Jul 18 21:07:57 EST 1994

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

Benoit_Hebert at iaf.uquebec.ca

Armand-Frappier Institute
Virology Research Center
Laval, Quebec, Canada    H7N 4Z3
phone: (514) 687-5010, ext. 4636
fax: (514) 686-5626

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