Boehringer DIG system experiences

[Georg Kroeger]

kferri at email.unc.edu (Karen Ferri) wrote:



>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
Hi Karen,

we use Boehringer DIG since many years successfully with Amersham's
Hybond N+ (charged membrane). We also use uncharged membranes
from time to time, but as blotting is easier and faster with charged
membranes (simply alkali-blotting and no baking or crosslinking
required) and as one can be sure that no nucleic acids will go
through, we prefer N+.

Cheers,

Georg

(no affiliation bla bla...)