Help! Direction of ligation
Keith.Leppard at warwick.ac.uk
Tue Sep 24 06:57:56 EST 1996
ialibova at ARTSCI.WUSTL.EDU ("Irina A. Libova") wrote:
> I am ligating an insert into a vector, using HindiIII site on both
> ends. And I need a specific orientation. But the problem is that on
> ALL of my clones I get the reverse orientation compared to the one I
> need, instead of getting a random 50:50 distribution of orientations.
> Has anybode had the same problem, and what can I do to change the
The orientation you want may be causing slow growth without being
completely lethal. Have you tried leaving the plates longer and
examining any small colonies that come up late? And/or culturing plates
at room temperature?
More information about the Methods