I have constructed one mutant having kanamycin cassette. For the growth comparison between wild type and mutant of same species, is it essential to add kanamycin antibiotic in liquid medium? In few papers, it was reported that if you not add kanamycin antibiotic in medium for the growth of mutant, kanamycin cassetted might get excluded by cells after 2 or 3 generation and behaves like wild type in growth curve.
Alejandro Martin
It depends on a lot of factors (including details of the cassette, how it is inserted, etc). In your case, I would run a trial curve of the mutant without kanamycin and compare cfu counts on plates with or without Kan at the end, to see if there is really a significant portion of the population that is losing the marker.
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