AflII restriction enzyme problems

[Bernard Heymann]

In article <D1vtK4.sE at ncifcrf.gov>, reming at ncifcrf.gov (Mary Remington)
wrote:

> Does anyone on the net have experience with this enzyme?  Does it cut and
> re-ligate reliably?  Any input appreciated.  Thanks, Mary

I've been using AflII a lot to screen for a particular insert in plasmids,
and it sometimes seem sensitive to too high salt concentrations in the
minipreps. I've only tried cloning AflII digested fragments once or twice
before, without success. The NEB catalogue says that the religation of
AflII cut fragments is low and one should use much larger amounts of
ligase.

Hope this helps.

-- 
Bernard Heymann
bheymann at bragg.bio.purdue.edu