Electroporation of COS: How to?

Ian A. York york at mbcrr.dfci.harvard.edu
Tue Sep 27 11:42:19 EST 1994


>In article <1994Sep27.160937.18429 at nomina.lu.se> Tomas Bratt 
><TOMAS.BRATT at MEDKEM.LU.SE> writes:
>In article <1994Sep27.161921.1 at icrf.icnet.uk> , g_watson at icrf.icnet.uk
>writes:
>>	I'm trying to electorporate some COS-7 cells with a vector containing
>>some immunoglobulin genes (well its two different vectors one for each
>>chain). 

>We have used 0.3 kV at 500 microF in PBS pH 7.4 in a cuvette with 4 mm
>gap with good results for both CHO and COS cells. However, we have not
>optimized the procedure for COS, only for CHO cells. Use15-30 microg
>plasmid. You have to be really fast to get the cells out on a dish after
>the electroporation or they will not survive.

	I am just about to do my first attempt at COS 7 electroporation.  
I got a protocol from The Guy Down The Hall, who has optimized it for our 
brand of COS7.  He used 0.8 ml cells at 3x10^6 cells/ml, 800 microF,"low 
ohms" and 200 V (yields 500 V/cm, it sayd here), in RPMI medium (no 
supplements).  He had used 5 ug of DNA but said that 10 would probably have 
been better.  
	Hope this helps.

Ian  
-- 
Ian York   (york at mbcrr.harvard.edu)
Dana-Farber Cancer Institute, 44 Binney St., Boston MA 02115
Phone (617)-632-4328     Fax  (617)-632-2627




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