Northern blotting analysis.

Knud Josefsen, Henrik Nielsen

The Bartholin Institute, Copenhagen University Hospital, Copenhagen, Denmark. knud@eln.dk

Methods in molecular biology (Clifton, N.J.) 2011

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Northern blotting analysis is a classical method for analysis of the size and steady-state level of a specific RNA in a complex sample. In short, the RNA is size-fractionated by gel electrophoresis and transferred by blotting onto a membrane to which the RNA is covalently bound. Then, the membrane is analysed by hybridization to one or more specific probes that are labelled for subsequent detection. Northern blotting is relatively simple to perform, inexpensive, and not plagued by artefacts. Recent developments of hybridization membranes and buffers have resulted in increased sensitivity closing the gap to the more laborious nuclease protection experiments.