undigestable genomic DNA

Warren Gallin wgallin at gpu.srv.ualberta.ca
Fri Feb 18 16:56:24 EST 2000


    At the risk of stating the obvious, different enzymes cut DNA from
different species to a different modal size.  I assume that this is partly due
to different site probabilities arising from differences in base composition,
but there are probably other general factors as well.    The only ways that I
know of to really tell whether the enzyme has worked or not are a) you have
done this digestion before on DNA from the same species and got a smaller modal
size or b) do a Southern blot with a single copy gene probe and see if you get
the unique pattern that you would expect.

Rafael Maldonado wrote:

> > Gregor Bucher wrote:
> >
> > > Hi southern-freaks!
> > >
> > > I want to do a southern with genomic DNA (insect). When I digest the DNA
> > > with Hind3 it looks fine. With EcoR1 and Xho however, there is quiete a
> > > bit high molecular DNA that obviously has not been digested (digesting 2
> > > hours or o/n does not change things). Does anybody have experience with
> > > genomic southerns?
>
> You could add spermidine, sometimes it is an improvement.
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