>>I'm want to find which method of transfection results in the highest
>percentage of cells taking up DNA. I want to do a transient transfection,
>followed by a western. At present my western signal is very weak if present
>at all. I think this is due to only a small number of cells being
>transfected. I am presently using chicken embryo fibroblasts.
>>Your thoughts on this would be helpful.
We've had good luck electroporating primary CEF's with RNA. I imagine you could
adapt these conditions for DNA:
0.2mm gap cuvette
0.5ml cells in PBS (we use RNase-free), 2x10e7/ml
BioRad GenePulser settings:
Program in Immunology
Washington University - St Louis
brett at borcim.wustl.edu