PCR deletion mutagenesis
Robert Horton
horton at cbs.umn.edu
Fri May 5 11:07:34 EST 1995
Fred Wagner (HEMA017 at UABDPO.DPO.UAB.EDU) wrote:
: Does anyone know of a protocol to make 25 to 100 bp deletions using PCR? I
: have seen various protocols for single site mutations but not for anything
: this large. Thanks. Fred Wagner
Large deletions are made the same way you would do recombination with PCR.
You amplify the two parts, using primers that add sequences so that the
parts have a small overlap. The parts are then joined by overlap extension
(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 biosci.cbs.umn.edu/(612) 624-3790
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