Big problems with cell-transfection

David L. Haviland, Ph.D. dhavilan at IMM2.IMM.UTH.TMC.EDU
Thu Nov 20 18:36:33 EST 1997


At 14:54 11/18/97 +0000, Tim Hatch wrote:
>In article <01bcf3a3$00827620$82019386 at schmidtdw.rz.ruhr-uni-bochum.de>,
>"Thorsten Schmidt" <Thorsten.Schmidt at rz.ruhr-uni-bochum.de> wrote:
>
>=A7I tried two different liposomes:
>=A7DOTAP (boehringer) and DAC-30 (Eurogentec).
>
>I've spent the last 2 years transfecting endothelial cells, and have been
>constantly frustrated by low efficiencies.
>
>DOTAP didn't work at all for me, (haven't tried DAC-30), but SuperFect
(Qiagen)
>has given efficiencies of around 40% (from FACS analysis of GFP
>transfectants.)

Tim:

I once needed to do transient transfections on HEK cells.  In short, I
found them resistant to most every means (DEAE, Electroporation, CaCl2, and
various lipids).=20

I was handed a sample of BMB's FuGene and it worked better than any of the
4 listed above.  However, at the same time, I also tried Clonetech's
CalPhos maximizer, and that worked *very* well.  In fact, if was about 2
fold better than the Fugene.    I'd give the CalPhos a try...

BTW - The conditions supplied (by this group) for HEK's  with
electroporation work well for making stable transfectants but I incurred
far too much mortality for transient work.

Hope this helps and no affiliation with BMB,
David


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 David L. Haviland, Ph.D.
 Asst. Prof. Immunology=20
 University of Texas - Houston, H.S.C.
 Institute of Molecular Medicine =20
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 Internet:"dhavilan at imm2.imm.uth.tmc.edu"
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