I transformed three plasmids into Top 10 (without F') selected by Amp, Tet and Cm. The colonies sometimes grow good overnight as normal sometimes grow really small and needs longer time. but when I start the culture from the colonies, it has problems. They always grow very slow, the overnight culture look transparent like nothing inside, but when I leave them in the room temperature one more day, they will give cloudy culture. I used this cloudy culture to start the larger culture and then induce by IPTG. I got the products which catalyzed by the proteins produced by the three plasmids.
also I store the cloudy culture into -80, and then I start them using fresh LB without any antibiotics, they still not grow overnight and need like one more day.
anyone has any suggestion? I can grow them normally? Thanks.