PCR mediated insert ligation
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?
Gene 77:61 (1989)
NAR 15:7351-67 (1988)
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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