Method: Electroporate COS-7

jstrassw at OPAL.TUFTS.EDU jstrassw at OPAL.TUFTS.EDU
Thu Nov 21 11:02:44 EST 1996


Hello!
After a LONG time trying to get high ewfficiency transfection of COS-7 
cells, I finally got 50% transiently transfected with electroporation.  
CaPo4 only gave a few percent as did many lipofection methods.

So, here it is:
3-5 million cells dont bother splitting berfore transfection
0.4 cm gap cuvettes, 1050 microfaradays 220 mV
10 micrograms of plasmid (Quiagen tip)
Do it at room temperature and in complete medium, no pre incubation, no  
post incubation. Dump the mess onto 100mm dishes with 10 ml of complete 
meduim.  Wait 6 hours or over night.  Change medium.  THen adday as you wish.

ps.  get a GFP reporter (like pGreenLantern or whatever) because you see 
your effeicincy in a few hours instead of a long B-gal stain.

Now, will someone post a decent transfection method for primary human 
fibroblasts?

John


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John Strasswimmer, MD,PhD Candidate    | Phone (617) 636 8396
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