C1R stable transfection

takedatu takedatu at med.kurume-u.ac.jp
Sun Nov 18 22:23:47 EST 2001


Dear flowers,
I've been trying to transfect the pIRESpuro vector to C1R(Bcell) cells,but
only gotten negative result. I decided the condition of electroporesion by
GFP expression. and got about 30% efficiency at the 48 hours after the
transfection.  started the selection after 24h or 48h or 72h with
1micro-g/ml of puromycin which was determined by C1R killing curve. after
2weeks or more, I got only dead cells in my flask. is it because of the rare
occurence of the  homologous reccombination?? Any protocols for getting the
better transfectant? thank you for your help. rocco


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