Controls in stable transfection experiments

Ned Mantei mantei at cell.biol.ethz.ch
Sun Feb 28 14:21:38 EST 1999


In article <7bbacq$45b$1 at mserv2.dl.ac.uk>,
wolfgang.schechinger at med.uni-tuebingen.de wrote:

 
> I'd like to ghather your opinions on appropriate controls for stable 
> transfected cell lines:

I doubt that you can get a transfected gene to express at a high enough
rate that there would be "stress of protein overexpression"  from
expression per se, as there can be in E. coli.

If you want to control for effects of random integration, you should
probably analyze a number of colonies from both the control and the
"+gene" transfections.

I would suggest using the empty vector as a control. If you use the
reverse orientation, isn't there the danger that the minus-strand RNA it
makes would suppress translation of the endogenous gene, with who knows
what effect?

-- 
Ned Mantei
Dept. of Cell Biology, Swiss Federal Institute of Technology
CH-8093 Zurich, Switzerland



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