Transient transfection of Cos-7 (electroporation v DEAE dextran)

George gyeung at ucla.edu
Wed Feb 19 21:44:11 EST 1997


In article <3309DBA6.7829 at sheffield.ac.uk>, K.Mulcahy at sheffield.ac.uk wrote:

> Hi,
> 
> does anyone have any experience with transient transfection of Cos-7 
> cells? I have attempted a DEAE-dextran/chloroquine based procedure but 
> only succeeded in transfecting 20-30% of the cells with a ß-gal reporter 
> plasmid (tested 24 hours post transfection). Does anybody else succeed 
> in obtaining efficiencies much higher than this (say 50-70%)?
> 
> Also, does anyone have any experience of using electroporation for the 
> transient transfection of Cos-7 cells? If so, could you please let me 
> know what electroporation conditions you use and what transfection 
> efficiencies you are able to obtain.
> 
> Many thanks,
> 
> Kevin Mulcahy

I have used the DEAE Dextran method, the calcium phosphate method and the
lipofectamine (BRL Gibco) method for COS cell transfection. Lipofectamine
seems to be the best but expensive. The calcium phosphate method is based
on the BBS-calcium phosphate method described in the Current Protocol (
the big red binders ), very cheap and work most of the time. Dextran is
the worst. I hope this help.



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