Trouble with growth of transformed cells
Posted 18 February 2011 - 10:50 PM
I am a Biotechnology student busy with a techniques course. We are doing cloning. Everything has been going well, but for some reason my cells transformed with my recombinant plasmid DNA, as well as those transformed with the ligation control, refuse to grow.
I know my cells are competent as my positive control and all the subsequent dilutions grew just fine. The plates were also made from the same batch of LB broth, amp, tet, IPTG and X-gal. I also have recombinants - did an agarose gel analysis (cloned 1200bp fragment into linearised pBS, EcoRI used as restriction enzyme)
I really don't know what is going on. Did the same procedure twice now, with no growth, blue or white, on those two plates.
Any suggestions will be sincerely appreciated
Posted 19 February 2011 - 06:29 AM
Posted 19 February 2011 - 09:14 AM
Posted 19 February 2011 - 11:25 AM
Posted 22 February 2011 - 11:08 PM
So, I managed to increase the efficiency and I got a few colonies! So yes, thanks a lot!