Stable cell transfection

Hendrie Kain hkain at nki.nl
Tue Jan 20 11:59:31 EST 1998


On 13 Jan 1998, James wrote:

> 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.
> 
> 
Why an tranfection if you can tranduce your cellines with an retrovirus.
In that case you will get a higher transduction efficiency.
Normally we will put the cells on antibiotics after 2 days of recovery.

PS. If the transfection doesn't work well.....try transduction

Hendrie




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