Login

Join for Free!
116885 members


TOPO PCR/Cloning

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

Moderator: BioTeam

TOPO PCR/Cloning

Postby 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.

Thanks!
coppcw08
Garter
Garter
 
Posts: 5
Joined: Sun Nov 04, 2012 8:07 pm

Postby JackBean » Wed Nov 14, 2012 7:35 pm

That depends only on your primers, to which position are they designed.
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
User avatar
JackBean
Inland Taipan
Inland Taipan
 
Posts: 5667
Joined: Mon Sep 14, 2009 7:12 pm


Return to Molecular Biology

Who is online

Users browsing this forum: No registered users and 1 guest

cron