Non-radioactive Southerns and Northerns
mattman at leland.stanford.edu
Fri Aug 4 01:17:00 EST 1995
Helen McBride <hmcbride at hlthsci.med.utah.edu> wrote:
> We have used the Boehrenger Genius sytem for non-radioactive
>Southerns with great success. The only real difference is that you need
>to use their membrane (positively charged) for it to work best. All the
>steps are the same except for different prehyb/hyb solutions. The
>detection is just like any other ECL system and exposure times are
>usually short (recommended time of 1 hr is usually just fine).
> I tried to use this system for Northerns and had a major problem-NO
>SIGNAL. I followed all their directions and used the recommended
>prehyb/hyb which is different from that used with Southerns and it still
>didn't work. I probed the same blot with a radioactive probe made from
>the same batch of cut DNA with no problem, so I don't know what went
>wrong. Others I have spoken with at our institution have also had no
>success using it for Northerns. I hope this helps.
>hmcbride at hlthsci.med.utah.edu
I and several other people I know have used the Genius system for
northerns according to the manufacturer's conditions and have gotten
wonderful results. Not that this really helps, but they really can
work. Only problem from my viewpoint is major deterioration of
results with each round of stripping.
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