shimshady, on Mar 22 2009, 05:35 PM, said:
I did a ligation and transformed into dh5a cells and grew at 37C. I checked the plates 14 hours into incubation, there were either no colonies grown at that time or i missed them. Anyways i left them in there because i forgot about them and remembered they were in there and went to dispose of them about 36 hours into incubation and there were some huge colonies on the plates (about 5-10 a piece). Some coloneis had the little colonies around them, im assuming were satellite colonies, and some didnt. My question is, are the colonies still good for being incubated that long. I will test to seee if they contain the appropriate plasmid later but i want to know if something bad happens to them for being incubated for so long, the ligation probably worked so i hope they are ok, at least i think it worked. I did have 50ug/ml carbenicillin on the plates and they were stored in 4C for about 10 days before use. thanks for you help.
I suspect that the colonies could be something else could be contamination. You say that you cant see any colonies after 14 hrs but did after 36 hours. Do a quick colony PCR to check for the plasmid. But really its alot of trouble. Just plate a new one.