Inserting 30 nt - which method? - help

user at host.edu
Wed Dec 25 18:15:14 EST 1996


Hi, everybody.

I'm trying to insert 24 to 30 nt into my new clone, i.e. insert a tag near
a stop codon to preserve 3'UTR for protein function studies. I'm looking
at FLAG which is 8aa long or c-myc - 10aa. Unfortunately, there is NO
convenient restriction site.
Which mutagenesis method would work well in this case? How long should the
hybridizing part of my oligo be on each side (15nt?)? How much can I loop
out? Anybody has experience with a problem of the kind?

If you know of a way of doing it please e-mail me or post.
Thanks a lot.

                        Yuri S.Oleynikov
         Albert Einstein College of Medicine.
         oleyniko at aecom.yu.edu



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