We are trying to make the stable clone of SW480 colon cancer after transfecting with micro-plasmid containing GFP. At the beginning of transfection, GFP-expressing cells are a lot but in subsequent subcultures, with selection antibiotic (neomycin) treatment the GFP-expressing cells are going down and we lose them. How can we keep them growing without losing the transfected cells?
Suggestions are well appreciated and thanks in advance...
Farhad
Hello!
Did you try different neomycin dilutions? How much time after transfection do you plate cells with neomycin? There is probably something to do by playing of these parameters.
Bests, Sandra
Dear Sandra,
Thanks for your response. We have generated kill curve for the cell lines used with different concentrations of neomycin and optimum dilution was used for the selection. We started antibiotic treatment after 2-3 days of transfection.
Thanks once again...
Farhad
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