Establishing stable cell lines...

Jump Jonny jonny_jump at yahoo.com
Wed Sep 13 19:02:45 EST 2000


Dear All

I am trying to establish stable mammalian cell lines
expressing a transfected gene by G418 selection in CHO
cells.

I treat the transfected cells with G418 @ 400ug/ml (I
assume this is this enough ?) but don't see extensive
cell death or gain much expression after 2 weeks
selection.

Could the problem be that I use Lipofectamine 2000 and
get so much DNA into the cells that too many of them
remain resistant to the G418 ?  Might a less efficient
transfection protocol like calcium phosphate work
better in the long run ?

Thanks in advance for any advice.

JJ

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