Stable transfection in suspension cells
Kamalian, Laleh
via methods%40net.bio.net
(by L.Kamalian from liverpool.ac.uk)
Fri Jan 18 17:49:02 EST 2008
Hi to everyone. I am a PhD student and new to this mail list. I have a problem I would like to ask. I am working with cells which grow in suspension. I have managed to get a good nock down in transient transfection using a plasmid vector and Lipofectamine as the transfection reagent. Now I am ready to perform the stable transfection. However I haven't got a clue how I will be able to separate the dead cells from the live ones in the selection process especially after the massive death happens. Furthermore, I don't know how I'm supposed to pick a single clone even if I manage to get the live ones. No one in our lab has ever transfected cells in suspension. The selection method I will use is antibody selection (G418) for which I have already started the optimization experiment in order to obtain the killing curve. I was wondering if any of you have worked with suspension cells. Could you please give me some help? Thanks
Laleh.
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