Fundamental Northern Blot question
kaghoram at garnet.acns.fsu.edu
Sun Jan 23 17:02:16 EST 2000
I have a couple of quick questions about Northern blotting. I use
GeneScreen Nylon membranes to capillary-blot polyA RNA.
Hyb with gene-probe1 -- 50 degC, wash 3-4 times at hyb temp; nice clean
Strip blot by boiling in 0.1 X SSPE; 1 % SDS for 15-20 min. Take care to
avoid drying; creases etc.
Hyb with gene-probe2 -- same conditions as first hyb.
Result: Blot covered with ugly spots. Even high-stringency washes don't
work. Heck, one time reboiling didn't take it off.
Haven't tried hybing with probe 2 first. Will try. But any of y'all have
any ideas why this may be?
Thanks a lot.
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