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transfection in eukaryotic cells - (Dec/15/2005 )

what is the principle behind transfection in eukaryotic cells ..mammalian cells
when we transfect a plasmid does it go into the nucleus or it remains in the cytoplasm

how can we do these transcfections..which are the most efficient ways and what are the differences between these ways
does anyone have answers to these questions
also can we co-transfect two plasmids using lipofectamine
what are the concentrations one can start with for luciferase reporter assays


check this guide:

Gives a good explanation for most stuff.
You can co-transfect 2 plasmids with any reagent, I've used fugene6 on U87 cells for 2 plasmids and lipofecatmine2000 for PCR-product and a cut vector (for homologuous recombination). Efficiency isn't that high of course, but it works. I've also done co-transfection on 293-T of 2 plasmids using calcium-phosphate, and this works also.

Apart from this, different cell lines require different reagents and conditions, so you better go the cell biology forum and ask there if anyone has experience with the cell line you're working with.