Do we need to keep puromycin in the culture medium during culture and induction of cell line?
We add puromycin (Final 1.0 µg/ml) into culture medium for isolated clone cell culture. We increase puromycin concentration to 1.5 µg/ml 3 days before transgene induction because higher puromycin concentrations help to enhance transgene expression ( Masui et al. Nucleic Acids Res., 2005 ). However, some types of ES cell differentiation seem to increase cellular sensitivity against puromycin. In this case, titration of puromycin concentration may be required.
