Anything and everything cloning: Go...
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Hi all,
I've been doing RF cloning for quite a while. I always worked with vectors of an final length of 6 kbp without any major issues. With 9 kbp I run into troubles. I end up without colonies. I'm pretty much sticking to the literature in my protocol. In the primer design, I must stick to a certain spot. So, the overhangs of the megaprimer a directly adjacent on the parental vector. I cannot leave out any bp in between. Could this also be an issue?
Any comment appreciated. Thx
Chris
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