Liner selection marker and stable transfectant - (Jun/06/2007 )

Hi everyone,

I'm trying to establish stable cell lines expressing a gene of interest in a plasmid encoding neomycin resistance.
Cause i' not lucky, my cells are naturally resistant to G418. So i want to cotransfect my plamid of interest with a plasmid encoding another selection marker. Do you think it's good approach?
Also, i'm wondering if someone used linear selection marker (sv40prom-Puromycin or Hygromycin- polyA form clontech) and get good results?


Thank's for your help!

Aurore

You could try to use 20:1 (interest:selection) plasmids for co-transfection. Linearize both plasmids may help. Once in the cell, the linear DNA join end to end, forming concatemers and integrated into the genome by chance.