Non-radioactive Southerns and Northerns

Helen McBride hmcbride at hlthsci.med.utah.edu
Thu Aug 3 17:54:44 EST 1995


     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



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