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G418 selection

G418 selection - Protocols and Methods Forum

G418 selection - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.

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Old 11-15-2005, 06:40 PM
Raheleh Masoudi
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Default G418 selection

Dear Sir/Madam

I have some monolayer HEK-293 transfected with Lipofectamin 2000 ( Invitrogen). 28 hours after transfection, I splitted the cells 1:8 and the following day I added different concentrations ( 50 microgram/ml 400 and 800 microgram/ml) of G418 for selection. Ater 7 days I could see there is difference between negative control ( transfected cells with no G418) and other wells in terms of confluency. After 9 days I changed the media and increased the amount o fG418 to 200, 500 and 1000 microgram/ml). Now I wonder how 293 colonies looks like. When I see the colony, how should I pick it up?

Sincerely yours,
Raheleh Masoudi
Graduate student

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