antibiotics concentration for ligated plasmid - (May/26/2005 )
hi....
i wanna know if the ligated plasmid got 2 different antibiotics..do we need to culture them (after transformation)in the exact concentration as we do before the ligation process?..lets say plasmid A need to be culture in 50ug/ml kanamysin.plasmid B=100ug/ml. if i ligate them both..what are the antibiotics concentration needed to be done?
Don't quite understand what you are doing. Do you merge two plasmids into one or transform two different plasmids in one transformation. Not a plasmid needs a certain concentration of antibiotics but the bacteria which receive the plasmid gets the antibiotic resistant marker from the plasmid and becomes resistant to antibiotics. So just stick to the concentration you normally use.
yup..im merged two plasmid into one and the ligated plasmid should have 2 antibiotics resistance..
hi you should try at least 100µg for good grow. The B plasmid confers a resistance to this level of antibiotic. And if A is expressed, don't care about it. Just take in mind that if you have B 100µg is ok. Moreover, that will kill bacterias clones that get only the self ligated A plasmid.