When do you starve cells after transfection?

aeschba at fmi.ch aeschba at fmi.ch
Thu Nov 3 11:45:24 EST 1994


Dear netters,

My question:
How quickly after transfection can you start to starve the cells. Will cells 
that are starved the same day that they were transfected still take the DNA 
correctly up and express it once you induce the cells?

I transfect cell lines with plasmids expressing the luciferase reporter gene 
under the control of various promoters. I starve the cells 24 to 48 hours after 
transfection with 0.5% FCS overnight (more than 16hours), then in the morning 0% 
FCS for 3 to 4 hours. At this point, I induce with different compounds. Analysis 
of luciferase is in intervals up to 24hours later.
I'd like to shorten now the 
time between starvation and induction, since quite often I take time points 
which are up to 96 hours after transfection. Also how do you correct for simple 
induction of metabolism and growth, which leads to more gene expression. I use 
SV40 promoter as a reference point, but I wonder if it really is a good 
"baseline".
Thanks for your answers.

Dr. Roger Aeschbacher
c/o CIBA
Pharma Research
Mauerstr. K 681-304
4002 Basel

E-mail: aeschba at fmi.ch




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