I am using the US02 cell line to perform an siRNA knockdown with an lipofection-based method (lipofection 3000). It has a puromycin selection marker. However, I am not sure what the best working concentration for a puromycin selection would be. Does anyone have any experience with this?
U2OS you meant? Article High Content Screening Using New U2OS Reporter Cell Models I...
In this article, the concentration of puromycin is 0.5 ug/mL.
Do note you should perform your own killing curve, due to expression difference between plasmid construction and heterogeneity between cell line passages.
You will have to do a kill curve using puromycin on the cells to determine the best concentration to use. Personally I do use (1ug/ml, 0.5ug/ml, 0.25ug/ml and 0.1ug/ml) when doing kill curve.
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