I transformed New England Biolabs High Efficiency E. Coli cells with a pBLUESCRIPT vector and my 1.2 kb insert. I got colonies that were 4x more abundant on the experimental LB-AMP plates than the control ligation plates, so it seems like some of the cells should have my circularized construct. But when I inoculated 10 colonies off the plate into liquid LB-AMP, none of the cells grew. To check if there was something wrong with the liquid media, I inoculated my media with E. Coli that was not from my plates, and it grew normally. The only culprits I can think of is that my cells did not take up the amp resistance vector, but grew anyway because ampicillan levels were not high enough on my plates, or that there is something wrong with the cells themselves. I also made sure to avoid picking satellite colonies.
Does anyone have any other ideas as to what could be going on?
I see that another poster is experiencing a similar problem.
Edited by Ceremony6, 02 February 2011 - 01:07 PM.