Hi there --
I'm hoping someone has an explanation (or even better, a solution) based
My student is trying to subclone a 4.1 kb Xba fragment into pCaSpeR-hs
(ca. 8.8 kb) in preparation for egg injections. The problem is that when
we ligate and transform bacteria, the resulting plasmids are either just
recircularized vector (no surprise there) or uniformly too small -- each
of these consists of a 4.1 kb fragment and a (roughly) 2.7 kb fragment
when re-cut with Xba. What's happened to the pCaSpeR-hs?
The insert has already been subcloned into another vector, so it seems
to be tractable. Has anyone ever run across something like this? We've
tried this several times, always with the same results. As you can
imagine, it's very frustrating (especially for my student).
Thanks for any advice.
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