Use this protocol if you have hybridization bags.
· 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
· 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.
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. 10 15 ml for prehybridization
b. 3.5 ml for hybridization
1. Prewarm buffer: Place the correct amount of DIG Easy Hyb in a sterile tube. Place