I am using gastric cancer cell line for my research, i used Lipofectamine 2000 for transfection, could any one recommend me the good concentration of G418 and Puromycin?
regards
Hi Farhan, I agree with Rachel that the appropriate concentrations depend on the cell lines you are working on. And a kill curve is usually recommended to determine the best working conc. However, if you would not like the trivials, I may advise that you search the published papers for your cell lines to get an idea, for example, to just Google it..
400 to 500 µg/mL would be a good starting concentration for G418 slection. (see attached G418 protocol with cell line dose differences)
Agreed with Dr Zhou, a kill curve would help.
Best,
Charles
Hi Farhan, I agree on all suggestions, I just want to provide you a nice template for your killing curve. See link:
Like Rachel, I would say that it is highly dependent on the cell line, but for most of colon cancer cell lines we use 1mg/ml of G418 and 1ug/ml of puromycin
Thank you all for nice and helpful suggestions, i will go through these steps..if i need further guidance will let you know.
bundles of thanks
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