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stabel Transfection of Jurkat cells


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#1 NiNaNu

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Posted 02 February 2009 - 03:01 AM

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

#2 scolix

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Posted 04 February 2009 - 03:25 AM

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.

#3 rkay447

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Posted 04 February 2009 - 05:04 AM

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.




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