Northern Blotting

Northern Blotting

Adapted from Molecular Cloning (Maniatis), Current Protocols in Molecular Biology, and J. Yost Lab (UMN)


Northern blotting allows detection of specific RNA sequences. RNA is fractionated by agarose gel electorphoresis, followed by transfer (blotting) to a membrane support, followed by hybridization with DNA or RNA probes.


Start=> This protocol requires isolated total RNA or poly(A)+ RNA.

  1. You must first run the RNA on a denaturing gel; a formaldehyde or glyoxal/DMSO gel.
  2. Next, you need to transfer the RNA from the gel to a solid support membrane, preferably nylon.
  3. Once, the membrane with the immobilized RNA is available, hybridization with the appropriate probe is required.