I am going to be using TOPO cloning for the first time. I am using the Invitrogen p-ET-102 Directional cloning kit.
My question is the following:
Will I be using my entire recombinant protein? It says that I need a blunt-end PCR product but I’m not sure if that mean’s I’ll have my product chopped along the way to create that blunt-end.
Anyone ever use this kit before? I had to add "CACC" to the 5′ of my forward primers as a recognition site but I don’t know if it uses my entire recombinant protein or not.
That depends only on your primers, to which position are they designed.