Striping and reprobing northern blots?
alansmith at students.wisc.edu
Thu Aug 16 09:58:03 EST 2001
I am looking for some advice on stripping and reprobing northern and
southern blots. I am trying to figure out the best method of stripping
blots to maintain a good signal and still strip the majority of old probe
from the blot. I have been using boiling 0.1% SDS to strip blots for some
time, but it seems to cause the northern blots to go bad after several
treatments (The sensitivity seems to decrease greatly after 3 strips. I
know there are numerous ways to chemically strip the probe from the blot for
example this works well for southerns (100mM NaOH, 10mM EDTA, 0.1% SDS) and
neutralizing the blot in 5x SSPE. Could anyone offer me some advice on how
to strip northern blots very efficiently and gently? I need to probe the
same northern blot as many as 12 times before checking the control probe.
Are there any solutions I should avoid for northern blots that are normally
used with southern blots i.e. NaOH?
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