Degenerate PCR - Sequencing?

Frank Richardson FRICHARD at biomed.med.yale.edu
Mon Mar 7 23:48:08 EST 1994


In <GERLACHJ.3.2D7B7997 at QUCDN.QueensU.CA> GERLACHJ at QUCDN.QueensU.CA writes:

> We have been doing PCR with degenerate primers (17mers, usually 256 fold) 
> and obtaining products in the 150-300 bp size. After doing restriction 
> digests to establish that we had produced some non-human products we are now 
> ready to sequence. We could, of course, clone and sequence in a conventional 
> fashion but I'm wondering what are the chances of doing sequencing using the 
> degenerate primers. Anyone tried this (or some other nice trick)? Thanx for 
> any sugguestions or help.

I have been doing PCR w/ degenerate primers as well and to date have not
encountered any problems using them to sequence as well - although I also use
the T7 and Sp6 sites on the vector as well.
virtually,
FR


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frichard at biomed.med.yale.edu (vms/vax system)              
"Life is not easy for any of us. But what of that?"
 - Marie Curie
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