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Transfecting CV1 cells

Bernard Murray bernard at elsie.nci.nih.gov
Tue Feb 4 00:07:53 EST 1997


In article <5d6c2f$2mci at news.doit.wisc.edu>, 
klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu says...
>
>In article <32F64107.51E3 at umich.edu>, Lourdes Rodgers <lrsdr at umich.edu> 
wrote:
>->Does anyone know what is the best or an efficient method of transfecting 
>->cv1 cells?  Has anyone tried calcium phosphate?
>->
>->Thanks for the help,
>->Lourdes Rodgers

>AFAIK, DEAE dextran is a classics for CV1. Electroporation will also work,
>that's for sure. 

Lourdes,
I've had efficiencies as high as 85% transfecting T25 flasks of CV-1
using the calcium phosphate method (can't get it as high using larger
vessels) as assessed by X-gal staining of pCMVbeta transfected cells.
You can expose the cells to the precipitate overnight and you don't
have to glycerol shock them.  The main thing is to have the cells
growing well before you plate them (and don't plate them at too high
a density).  Stable clones can easily be selected.
	Lipofection (Gibco's LipofectAMINE) works just as well but
costs more, of course.  Again, you can get stable clones easily.
	Following up on what Dima says I've also heard good things about
DEAE/dextran but it is much more difficult (?impossible) to generate
stable clones with that method.  I have no personal experience with
electroporation of this cell line (I reserve that for transfection
resistant lines).
	All the very best,
		Bernard

Bernard Murray, Ph.D.
bernard at elsie.nci.nih.gov (National Cancer Institute, NIH, Bethesda MD, USA)




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