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selection of stable transfectants with ampicillin - (Sep/24/2006 )

Dear All
I have transfected a human cancer cell line with a plasmid that has an ampicillin resistance gene, WHAT is the concentration of ampicillin should I start with? is there any important notes to consider during the selection process?


I might be silly here, but Amp selection shouldn't work on an eukaryote. Correctly me if I am wrong but eukaryotes don't have cell wall synthesis for ampicilin to block. Wasn't that the whole magic bullet idea when penicilins were first dicovered?

Try something else like G418, the resistence gene to this also gives bacteria resistence to Kanamycin


perneseblue is right.
check on your plasmid map. the ampicillin resistance gene is for selection of the bacteria. There should be an other resistance gene with the eucaryote promoter.


sorry for the terrible mistake


QUOTE (yobou @ Sep 25 2006, 11:43 AM)
sorry for the terrible mistake

no problem.

by the way, you should treat non transfected cells with various concentration of antibiotics to determine which concentration is the best for your cells. some cells can be more or less sensitive to antibiotics.