Stable cell transfection

James jsg at mole.bio.cam.ac.uk
Tue Jan 13 08:12:01 EST 1998


A couple o' questions :

1. Is it absolutely necessary to linearise the plasmid before
transfection? My cell lines cannot support plasmid replication, so
integration is required.

2. How long after transfection should one wait before adding antibiotic to
the culture medium? I'm using hygromycin B, and transfecting by
electroporation.
One of the cell lines is adherent (EL4), the other grows in suspension (P815).

Any advice welcome
James.



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