bernard at elsie.nci.nih.gov
Mon Sep 16 17:05:38 EST 1996
In article <Pine.SOL.3.95.960913091324.5615L-100000 at suma3.reading.ac.uk>,
saspeyer at reading.ac.uk says...
>There's just one problem with the pGEM-T vector - it no longer exists!
>I had designed all my primers around it, but when I 'phoned them up to
>order some more yesterday I was told that it had been withdrawn due to a
>patent dispute! To everybody else who is a current user of pGEM-T...
>jeez, just run for the hills, that's all I can think of.
Nah, just run for your (approved) thermocycler. Since pGEM-T is
just pGEM-5Zf(+) cut with EcoRV and T-tailed you can make as much
as you want (I think there are procedures in the FAQ based on Taq/dTTP
or other enzymes and ddTTP).
If you are lucky and your insert doesn't have an EcoRV site
you can do a blunt ligation in the presence of EcoRV (much the same
as Stratagene's pCR-Script and their rarer cutting SrfI).
Clone to your heart's content...
Bernard Murray, Ph.D.
bernard at elsie.nci.nih.gov (National Cancer Institute, NIH, Bethesda MD, USA)
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