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Not Telling
Hi all,
I've performed some degrees of cloning before but I never really understood how you could check whether or not your insert is 'in-frame'?
Previously when I cloned, I would get a blunt PCR product which I did a blunt end ligation into a TOPO vector. Then, I used a restriction enzyme
that cut my insert ouf of the TOPO vector, clipped another vector, and ligated that together. Professors have always told me to check to make sure that my insert goes in 'in-frame', but how would I do that in terms of designing my primer?
If I use the digest-ligation method like I mentioned above, would you assume that it's going in frame?
