TOPO PCR/Cloning

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TOPO PCR/Cloning

Post by coppcw08 » Wed Nov 14, 2012 5:59 pm

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.


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Inland Taipan
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Post by JackBean » Wed Nov 14, 2012 7:35 pm

That depends only on your primers, to which position are they designed.

Cis or trans? That's what matters.

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