Transfection

Bernard P. Murray, PhD bpmurray*STUFFER* at socrates.ucsf.edu
Fri Aug 28 20:07:32 EST 1998


In article <6s6tfh$i50$1 at wnnews.sci.kun.nl>, "Gerrit Bouw"
<gbouw at sci.kun.nl> wrote:

> Hi there,
> 
> I am in desperate need for a transfection protocol which gives up to 60 or
> 70% transfection
> efficiency in CV-1 and AtT20 cell lines. Are there such protocols anyway? I
> would be
> very happy to test different protocols on my cell lines...!
> Thanx..
> Gerrit

I have seen such efficiencies with CV-1 cells using standard CaPO4
or Gibco's LipofectAMINE.  Optimise for your particular circumstance
but probably the key thing is to use cells that have not been
passaged very often.  The efficiency drops fairly precipitously with
increasing passaging but the cells can be made more "competent" by
freezing them as normal and then starting the cultures anew.
The cells are fairly hardy so you can expose them to CaPO4 or
LipofectAMINE overnight.
     I hope that this helps,
          Bernard
-- 
Bernard P. Murray, PhD
Dept. Cell. Mol. Pharmacol., UCSF, San Francisco, USA



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