Northern Blotting despair!

punk at leleand.stanford.edu punk at leleand.stanford.edu
Thu Aug 14 13:35:29 EST 1997


Hi what is the problem no signal, high background. You  should be able
to detect low abundent message with total RNA  depending on how low
abundent. You can also use poly A selected RNA  we use dynabeads for
this very easy and fast. If no signal check the transfer using methylene
blue staining of the membrane, procedure found in the red book.  ALso
try to probe blot with actin or something more abundent to test hyb and
prob production procedures. Besure  you clean up you probe to get rid of
unincorporrated nucleotides.  I can't be any more specific unless you
describe you problem in more detail seeya gregg



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