stable transfection of balb/c 3T3

D.K. no.email at thanks.to.spam.net
Wed Mar 10 11:32:51 EST 2004


In article <20040310041656.31928.qmail at ww02.hostica.com>, fionsin at hkem.com wrote:
>Dear all,
>
>I'm going to do a stable transfection on balb/c 3T3 by using electroporation.
> Now, I'm testing the amount of G418 needed to kill the cells. I tried G418
> concentration ranged from 200ug/ml to 1000ug/ml but the cells do not start to
> die (It's already the 10th day after G418 addition). I maintain the cells in a
> density of 50% full and the medium(with drug) is refreshed every 2 days. Can
> anyone help me?

3T3 I worked with in the past were dying at 0.6 mg/ml. Do you go 
by G418 dry weight or by the active comopund content? Commercial
G418 is usually only about ~ 70% pure. Gibco always indicates
the exact value on the bottle. 

DK



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