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Northern Blot Use this protocol if you have hybridization bottles.
 

Summary: 1
Northern Blot
Use this protocol if you have hybridization bottles.
Hybridization of DIG labeled (non-radioactive) DNA probe
to a nylon membrane blotted with RNA.
Detection of the probe with Anti-DIG-AP antibody and
chemiluminescent AP-substrate.
Stripping the DNA probes off the membrane.
IMPORTANT: Once you have started this set of protocols, never let the membrane dry at
any point (not even after it has been stripped). If the membrane is allowed to dry, it cannot
be stripped. If handled properly, the membrane can be stripped and reprobed up to 14
times.
Hybridization
Determine how much DIG Easy Hyb you will need for the procedures below. You will
need enough buffer to completely cover the membrane during incubations. For 100cm2
of membrane, you will need:
a. 20 ml for prehybridization
b. 5 ml for hybridization
I. Prehybridization
1. Prewarm buffer: Place the correct amount of DIG Easy Hyb in a sterile tube. Place

  

Source: Andres, Andrew - School of Life Sciences, University of Nevada at Las Vegas

 

Collections: Biology and Medicine