Negative bands on Southern/Northern

Frank O. Fackelmayer fof1 at chclu.chemie.uni-konstanz.de
Tue Jul 1 05:20:20 EST 1997


In article <5odejh$fd0 at scotsman.ed.ac.uk>, pmcclive at holyrood.ed.ac.uk
(Peter McClive) wrote:

> We've recently had a run of blots which have a very high background
> except for regions with high DNA or RNA (markers or 18S/28S) which
> appear white.
> 
> One pair of blots generated from the same piece of membrane and
> hybridised in the same bottle gave a normal result and a negative
> result.  The only thing that differed was that they were blotted at
> different times.  Could the negative bands be due to excess salt left on
> the membrane after DNA/RNA transfer from the gel?

Hi Peter,
I had this problem some years ago, and I found that the reason was
insufficient blocking of the membrane. Try to find an error in this
direction...

Hope this helps,
Frank



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