My purpose is to generate stable cell line and I am selecting my cells with puromycin. My question is " Is it ok if I add puromycin in my working media (DMEM +10% FBS+ 1% pen/Strep + Fungizone) or I need to add it to DMEM+10% FBS (no antibiotics or funzigone)
If you your selection marker for stable construct is puromycin you need to maintain your cells in puromycin containing media (DMEM+1%P/S+10% FBS+ Puromycin -recommended conc is 5 ug/ml). However, I am not sure why you are adding fungizone in your media. Fungizone is only used whenever there is fungal contamination. Cells should not be maintained in Fungizone.
Thank you so much for your answer. I ma using fungizone because we are having some problem with fungal contamination in our lab.Once I get my single clone then I will grow and maintain that clone in a Media lacking Fungizone.
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