slot-blot

Raymond Dalgleish ray at le.ac.uk
Fri Jun 18 03:16:42 EST 2004


If you HAVE been using the nylon and not the backing paper, try the
following. Nylon membranes are very difficult to wet if there is "high salt"
in the solution. Try filtering some "low-salt" solution through the membrane
prior to applying the DNA samples. I find that 3 X SSC works fine for
pre-wetting nylon filters.

Raymond Dalgleish


"Dirk Müller" <Kraweel at web.de> wrote in message
news:3vh2d0lu1i2fjreeg0p2uoo675cmsk23c7 at 4ax.com...
> A stupid question but my students do this most of the time that
> filtrations do not work: Are you sure that you use the nylonmembrane
> and not the paper that is on one side of the membrane for cover?
> regards, Dirk...
> On 16 Jun 2004 13:38:41 +0100, marius.banholzer at uniklinik-freiburg.de
> (Marius Banholzer) wrote:
>
> >I´m doing DNA-slot-blots. The DNA is denatured by cooking with NaOH for
> >3 minutes at 95 degrees. For blotting a Nylon-membrane is used. The
> >problem is, that the probes are not sucked through the membrane. Even
> >the washing buffer won´t go through.
> >The vacuum is ok. Please help!
> >
> >---
>





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