Hi, I' am trying to make stable cell line using plasmid transfection.
the plasmid are available for neomycin selection.
I want to know the appropriate G418 concentration and time forselection about Mlg cell line. Mlg cell line is mouse lung fibroblast cell line.
and also I wonder the concentration and time of selection about mouse primary lung fibroblast.
Thank you
Many factors can impact selection in culture. The best advice is to perform a kill-curve using the G418 in your media and cell lines. Typical concentrations are 0.5-1mg/mL of G418. I would start at 2 -4 mg/mL and dilute down several fold, including a no selection control. You want to find the concentration that inhibits/kills cells within a few days. Stable transfections take weeks to sufficiently select and expand. Once expanded, you can maintain the cells at a lower concentration (e.g. 0.2 - 0.4 mg/mL).
okay. thank you very much.
following your recommendation, I am going to try make a kill curve.
thank you.
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