Selcetion of WISH cells with G418

Frank O. Fackelmayer Frank at Fackelmayer.de
Mon Apr 23 11:29:17 EST 2001


Hi Wolfgang,
I have no idea about the WISH cells, but 100ug/ml is very low. I don´t
think it will be sufficient to kill the non-transfected cells. Most
human cells I have used and heard of can stand at least 500ug/ml, and if
I don´t have an idea about the maximum amount the cells can stand, I use
this concentration. Actually most stable transfectants easily stand
800ug/ml, and I have the impression that it only makes economic (but not
scientific) sense to set up a kill curve to find the lowest dose of G418
that is sufficient to kill non-transfected cells. As killing of
"non-stables" with G418 usually takes up to two weeks, you will probably
not be happy with the "wait and see" strategy.

Frank




Wolfgang Schechinger wrote:
> 
> Hi all,
> 
> thanks al lot for theresponses on rice germination!
> 
> Has anyone already tried so select WISH cells (a human amnion
> derived cell line) for stable transfectants using G418? I would
> like to selcet some stable clones from a (actually transient)
> transfected dish and don't have a kill cureve for these cells
> yet. What concentration range might be suitable?
> 
> By now, I'll start with 100ug/ml to see if some clones survive
> and increase the dosage if there are too much..
> 
> All input is welcome,
> 
> Wolfgang




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