Boehringer DIG system experiences

Mike Challen mike.challen at hri.ac.uk
Mon Feb 3 04:26:05 EST 1997


Hi Karen 

We've been using DIG almost since it came on the market and have tried all
sort's of different membranes. Although there are subtle differences in
background and sensitivity, they all seem to work.   
We have most often used Hybond-N (without charge) but,  partly promted by
recent structural changes in Amershams membranes, we have switched to the
positively charged membrane marketed by Boehringer.
Any way, the straight answer to your question is;  No you do not have to
use a positively charged membrane.  Try what you have it will probably work
fine.

best wishes
Mike

Karen Ferri <kferri at email.unc.edu> wrote in article
<5d11p8$jap$1 at newz.oit.unc.edu>...
> 
> 
> HI everyone!
> 
> I am currently trying Boehringer's DIG Labeling and Detection kit b/c I'd
> like for our lab to switch over (me, at least) to non-radioactive
probing.
> 
> The instructions call for positively charged nylon membranes.  The one we
> have in lab is by Amersham and I imagine it is uncharged, since it didn't
> say anything on the box or instructions.
> 
> I did my control labeled DNA filter and the spots are black.  This was on
> the Amersham filter.  SO is a positively charged filter truly needed? 
I'd
> like to find out others' experiences before I go through the trouble of
> preparing a membrane with my experimental DNA
> 
> TIA,
> 
> Karen
> 



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