3 replies to this topic

[hogthehedge]

Hi everyone,

Do you know which antibiotic to use to select mammalian cells expressing a plasmid with Ampicillin resistance?
I use Geneticin for Kanamycin-resistance conferring plasmids. Would the same work?

Regards,
HogtheHedge

[leelee]

I don't think there is one.

No, the amp resistance gene will not confer protection against geneticin.

[hogthehedge]

Thanks for your reply, Leelee.
I have been given plasmids without maps or sequences. I know they confer resistance to amp, but I don't know what to select with once I transfect into mammalian cells. I am trying to make stable cell lines. If anybody has any idea, please do let me know. Thank you!

[bob1]

Amp does not work at all as a selective agent for eukaryotic cells; it acts against the cell wall of bacteria.

If you do not have the sequence, do you have a name for the plasmid?  If so, a lot of the backbone details are available in various places on the web.  It may also be that the plasmid you have is only good for transient transfection or bacterial expression.