stable cell lines

Muralikrishna Vemula vemula at helix.mgh.harvard.edu
Thu Mar 7 14:00:42 EST 2002


Hello,

I am working with HepG2-C3A hepatocyte cell line. My goal is to transfect
these cells with GFP reporter plasmids and establish stable cell lines.
The plasmids have Neomycin resistance gene in them. Can somebody please
help me with these queries?

1. Does anybody have a set protocol that describes the procedure starting
from trancfecting cells to performing gene expression studies using stably
transfected cells?

2. Do the cells usually have a different generation time after stable
transfection?

3. Do you have to screen for clones that behave as close as the wild type?
If so, how many do you have to screen?
 
4. How long does this whole procedure take? 


Cheers,
Murali Vemula

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