Hybond Background in Southerns - HELP

W R Bennett bspwrb at bath.ac.uk
Thu Sep 11 12:35:18 EST 1997

In the referenced article, z2154398 at student.unsw.edu.net (tom) writes:
>I'm using Hybond N (not N+) for genomic Southerns and am getting a high
>background even with Church buffer.
>Any suggestions for alternative buffers, membranes, or procedures?
We use Hybond-N (not N+) for genomic Southerns and after switching to 
Church, our background problem vanished.  Are you sure you're using enough 
volume of Church?  Amersham recommend 125ul/cm2 of membrane, and we use 
30ml Church per 20x20cm filter.  Also, we chuck away the pre-hyb and add 
30ml fresh Church buffer with the probe - the new Hybond-N soaks up 
buffer like a sponge.


Bill Bennett
bspwrb at bath.ac.uk

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