HELP with stable transfection - (Mar/19/2007 )

[b]i want to do stable transfection using liver cancer cells and exactly H7402 cell line .i am trying to find the right protocol to do the stable transfection. can somebody help me please !

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well i'ts an adherent cell line ? a suspension one ?
did you buy it or get it from an other lab? in these case they should mention protocol.
You can trry the lipofectamine or calcium phosphate transfection methods... (lipo is more efficent but caPO4 is cheaper)

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stable transfection often utilizes an antibiotics resistant factor mostly on the same vector as the gene of your protein of interest; e.g. a standard protocol for kanamycin/neomycin resistance appear to be by G418; selection may occur between 0.35 up to 1 mg/ml

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Hi,
Thank you so much for your reply! 'The Bearer' , and fred 33 (j ai prefere de te dire merci en francais )
i am very happy to find scientists that can help me
Ok. the cells that i am using is an adherent cell lines. I transfected a plasmid that cantains my gene of interest (VEGF) USING THE ELECTROPORATION method. For the seleciton, i used the G 418. After one week , all the cells died and i havn t get any transfected cell. until now, i don t know where is the problem ?

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