site directed mutagenesis kits

"Miss. V.M. Hodges", by way of florigen at Florida Genetics Corporation vhodges at
Thu Jul 25 11:50:51 EST 1996

Miss Hodges writes:

"Can anyone recommend a kit for doing site-directed mutagenesis - yes I
know its possible to do it without a kit  ( and I've done so in the
past) but I read about a kit somewhere and it seemed to be fast and
efficient . any suggestions would be welcomed (preferably from companies
supplying in the UK).

Vivien,  We have used a double stranded template protocol offered by
Stratagene (QuickChange) with good success.  All you need is Pfu, Dpn1
(backordered, we got ours from Boehringer), and two complementary oligo's
that flank the site.  The reactions and transformations can be carried out
in one day, and out of 8 (2 sets of 4) sent for sequencing, 75% contained
the mutation.  Be sure to check Pfu fidelity.  Good luck!


Wm. McKendree, Jr., PhD
Florida Genetics Corp.
Alachua, FL   32615

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