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Methods Enzymol. 2013;530:301-13. doi: 10.1016/B978-0-12-420037-1.00016-6.

Analysis of RNA by analytical polyacrylamide gel electrophoresis.

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1
Stanford University School of Medicine, Stanford, CA, USA.

Abstract

Polyacrylamide gel electrophoresis (PAGE) is a powerful tool for analyzing RNA samples. Denaturing PAGE provides information on the sample composition and structural integrity of the individual RNA species. Nondenaturing gel electrophoresis allows separation of the conformers and alternatively folded RNA species. It also can be used to resolve RNA protein complexes and to detect RNA complex formation by analyzing changes in the electrophoretic mobility of the RNA. RNA can be visualized within gels by different methods depending on the nature of the detection reagent. RNA molecules can be stained with various dyes, including toluidine blue, SYBR green, and ethidium bromide. Radioactively labeled RNA molecules are visualized by autoradiography, and fluorescently labeled RNA molecules can be observed with a fluorescence scanner. Generally, gels between 0.4 and 1.5mm thick are used for analytical PAGE. Gels thinner than 1mm are fragile and thus usually are not stained but rather are used for radiolabeled RNA. The gels are dried and the radiolabeled RNA is visualized by autoradiography.

KEYWORDS:

Analytical polyacrylamide gel electrophoresis; Gel preparation; Polyacrylamide meshes; RNA analysis; RNA visualization

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