ligation and transformation in DH5A
Posted 22 April 2010 - 05:29 AM
I am trying to put a PCR product (850bp) in the vector pet20b. I am doing the ligation at 16 degrees O/N and then I transform DH5A competent cells and plate them in plates with LB and ampicillin. Finally, I get no colonies or just one colony without the insert.
Do you have any suggestions? Thanks a lot!!!!
P.S. the self-ligation works correctly.
Posted 22 April 2010 - 02:29 PM
Posted 22 April 2010 - 08:53 PM
Posted 22 April 2010 - 11:47 PM
yes, that's exactly the problem! any suggestions or options????