PCR Replacement/Knockout

Richard John Brennan brennan at mbcrr.harvard.edu
Thu Jun 22 15:23:24 EST 1995


Mike Dalrymple (dalrymple at pplros.demon.co.uk) wrote:
: Hello,

: we have dabbled in the PCR knockout/replacement approach to yeast genome
: alteration
: without much success. Postings on this topic appeared some months ago and
: were
: helpful, but I've been told that current methodologies work EXCEPTIONALLY
: well. Could anyone update me on the particular parameters which lead to
: success
: with this technique. I understand that a paper on the technique has been
: published,
: is anyone familiar with it?

: Many thanks

: Mike Dalrymple
: PPL Therapeutics Ltd
: Edinburgh

There's a paper in press in NAR by Manivasakam, Schiestl and Weber
regarding length of homology required for targeted integration,
using PCR generated fragments. You might be able to get a preprint
for latest on transformation procedures etc. You can email Robert
Schiestl (schiestl at mbcrr.harvard.edu) or Mani (mani at mbcrr.....).

Rich.




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