I performed directional cloning of a gene into a vector.
I was wondering if you need to de-phoshorylate the ends even with directional cloning where the cut vector does not have compatible ends?
It would seem possible that a cut vector could ligate with another cut vector (not self ligate), so it would be a good idea to de-phoshorylate the ends when doing directional cloning?
What are peoples opinion? I just screen about 10 colonies and they are all negative, so there must be some form of re-ligation happening.