Co-transfection for stable cell lines - mammalian - Need to know the ratio of the plasmids that need to be used (Mar/08/2006 )

Hi Cell Biologists

I am doing a co-transfection to make a stable cell line. My plasmid with gene insert doesnt have a selection marker. So, I am co-transfecting with a vector with antibiotic selection marker.

I would request any of you to help me to figure out the concentrations or ratio of the two plasmids that has to be used for optimal transfection and to get maximum positive colonies expressing my gene of interest.

Aucklander
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Just make the ratio quite high, I know of some who had been quite succesfull in going 6:1, but I don't think there is a generall rule for this.

hi,
for good cotransfection efficiency, use 10: 1 ratio of your gene of interest to the selection marker plasmid.
it worked good for me...
good luck
syc06