Electroporation condition for Jurkat cells

Alan Lau AL18 at le.ac.uk
Tue Sep 10 10:50:57 EST 1996


In article <50gn2k$dhb at news.Belgium.EU.net>, r.carreer at eurogentec.be
(carreer) wrote:

> Conditions :
> 
> 
> 5 million exponential growing cells 
> spin down
> resuspend in 250 microliter of culture medium (not DMEM) RPMI is okay
> with 10% FCS
> place cells in cuvette of 4 mm
> Keep on ice
> 
> add 20 microgramme of plasmid
> 
> Perform Electroporation
> 250 Volt
> Capicity : 1200 microFarad
> shunt : infinite
> half time : about 45 to 55 mseconds
> 
> Place directly electroporated cells in the corresponding culture
> medium at 37°C


I use an electroporation method almost exactly the same as this and it
works reliably.  However, I cannot get electroporation efficiencies
greater than around 15-20% of the total surviving cells.

Does anyone have any comments, suggestions or alternative methods (e.g.
lipofection) to improve this efficiency?

Thanks in advance,

Alan.

-- 
al18 at le.ac.uk
Dept. Microbiology and Immunology
University of Leicester
LE1 9HN



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