I've transfected HEK293-T cells with pCI as a mammalian expression vector containing neomycin gene as a selectable marker. I need to know the most efficient concentration of G418 for cell selection.
Thank you all!
293T are 293 cells transduced with the large T antigen to boost expression of SV40-mediated expression and have a Neomycin resistance. G418 does not work to select transfected cells. Use another resistance gene / antibiotic combination or use the parental 293 cells.
I have successfully used 0.4 mg/ml of G418 for clonal selection. I'm not sure if this is the optimal concentration but it works.
293T are 293 cells transduced with the large T antigen to boost expression of SV40-mediated expression and have a Neomycin resistance. G418 does not work to select transfected cells. Use another resistance gene / antibiotic combination or use the parental 293 cells.
Michel Ouellet is correct. G418 will not work in 293T, but works well in 293 at 0.4 mg/ml as Elizabeth Johnstone suggested.
If you plan to stick to 293T cells and need a different antibiotic marker, I suggest using puromycin. It doesn't take as long to kill off untransfected cells.
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