pCaSpeR-hs cloning problem

Chris Jones 6kuritaro at earthlink.net
Sun Apr 6 21:07:57 EST 2003


Hi there --

I'm hoping someone has an explanation (or even better, a solution) based 
on experience:

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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