I am using pCR T7/NT-TOPO expression vector for expressing a plant gene. As a negative control I have to have empty vector expression. I have read somewhere that linear empty TOPO vectors cannot be transformed since they have topoisomerase at the cloning site. How can I then prepare E.coli cells harboring empty TOPO vector?
If I get such cells prepared by some means, can the isolated TOPO plasmid be digested in some way and used as a expression vector for ligation using ligase enzyme?
Edited by ram, 02 January 2010 - 02:13 AM.