Northern Blot Hybridization

Authored by: Kirakosyan Ara , Peter B. Kaufman , Tsai Chung-Jui , Leland J. Cseke

Handbook of Molecular and Cellular Methods in Biology and Medicine

Print publication date:  December  2011
Online publication date:  April  2016

Print ISBN: 9781420069389
eBook ISBN: 9781439881958
Adobe ISBN:

10.1201/b11351-14

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Abstract

Northern blot hybridization or RNA blotting is an RNA analysis procedure in which the size and amount of specific mRNA molecules in total RNAs or poly(A)+RNAs can be determined. 1 6 It basically consists of RNA/DNA or sense RNA and antisense RNA hybridization. RNA molecules are separated according to their sizes by electrophoresis on a gel under denaturing conditions, transferred to a nitrocellulose or nylon membrane, and fixed to the membrane. The blotted membrane is hybridized with a specific probe, and the mRNA of interest is then visualized by autoradiography or non-isotopic detecting methods (Figure 11.1).

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