I was wondering if anyone could help me. I have tried to clone 240 bp PCR product cutting with NdeI/XhoI (restriction site in the primer) into pET23a+ cut with the same enzyme for a long time. There was no positive clone after many times cloning. After changing the host strain to K12, there were clones and then I extracted the plasmid with 5 PRIME kit and double digested with NdeI/XhoI. After running the gel, plasmid band was found but no or empty band in the digested plasmid. The control pET23a cut with NdeI/XhoI showed correct band. These showed no error in water, RE , buffer or BSA). Please help me to clarify this.
Thanks in advance
Edited by Pig, 11 August 2010 - 01:53 AM.