PCR mediated insert ligation

Robert Horton horton at molbio.cbs.umn.edu
Fri Feb 18 15:28:38 EST 1994


douglas l feinstein (dlfeins at cumc.cornell.edu) wrote:
: We have a situation where we have cloned a 4 kb cDNA  in 4 separate
: pieces, and now want to stich it together to make a functional molecule.
: The inserts only overlap by  50 to 100 bp, so it is hard to find unique
: and overlapping restriction sites. I would think there's a way to use PCR
: to do this. Any ideas?
:

See
Gene 77:61 (1989) 
NAR 15:7351-67 (1988)
BioTechniques 8:528-535

Directed Mutagenesis: A Practical Approach (McPherson,M.J>, ed)
IRL, Oxford, pp217-47 (1991)

.. and many others. Look for "PCR recombination" or "overlap extension"
or "gene SOEing".


Bob Horton (Ph.D.!)   /\ "Crash programs fail because of the theory that
U. of Minnesota, CBS  || with nine women pregnant you get a baby a month" 
1479 Gortner Ave.    /||\   -Werner von Braun.  Disclaimer:"Bob who?"
St. Paul, MN 55108    ^^   horton at molbio.cbs.umn.edu/(612) 624-3790



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