I am trying to make KO in Huh7 cell line but after transduction even at 5ug/ml concentration Huh7-Ctrl cell haven't died. Could someone please recommend me the working concentration of Puromycin for Huh7?
What kind of puromycin are you using? In my lab, a student had a problem in his experiments because he used puromycin aminonucleoside which is not recomended for selection. To give you an idea, I tried puromycin selection with dermal fribroblasts (1,5 ug/ml), endothelium (0,5 ug/ml) and cybrids (1 ug/ml) and it worked pretty well.