stabel Transfection of Jurkat cells - G418 Selection (Feb/02/2009 )

Hi everybody,

I tried a stable Transfektion of Jurkat cells via elektroporation. Unfortunately non of the cells died, not even in the mock transfected control.

No I tried a titration of the G418 but even at 5 mg/ml non of the Jurkats died, they grow and replicate very nicely....

So is it possible, that I use a Jurkat clone, that has already a G418 resistence? Something like in the 293T cells?

I can't find anything liek that in the www, perhabs you can help me.


Cheers,

Nina

Try making fresh G418 from powder and try the concentration curve for G418 again.

Or it may be that the 293 is already selected using G418.

I agree with Scolix. There is certainly something wrong with the G418. The mock transfected are supposed to be a positive control for cell death so if they didn't die, the drug didn't work. There are a number of papers using this cell line and this drug for stable cell selection so this cell line is sensitive.