G418 selection

[Raheleh Masoudi]

        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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