Northern Blotting

Northern BlottingAdapted from Molecular Cloning (Maniatis), Current Protocols in Molecular Biology, and J. Yost Lab (UMN) Objective: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.

Procedures: Start=> This protocol requires isolated total RNA or poly(A)+ RNA. You must first run the RNA on a denaturing gel; a formaldehyde or glyoxal/DMSO gel . Next, you need to transfer the RNA from the gel to a solid support membrane, preferably nylon. Once, the membrane with the immobilized RNA is available, hybridization with the appropriate probe is required.