Horizontal Gel Electrophoresis for Enhanced Detection of Protein-RNA Complexes

Overview

Journal J Vis Exp

Specialties Biology
General Medicine

Date 2017 Aug 9

PMID 28784977

Citations 5

Authors

Megan E Dowdle

Susanne Blaser Imboden

Sookhee Park

Sean P Ryder

Michael D Sheets

Affiliations

Soon will be listed here.

Abstract

Native polyacrylamide gel electrophoresis is a fundamental tool of molecular biology that has been used extensively for the biochemical analysis of RNA-protein interactions. These interactions have been traditionally analyzed with polyacrylamide gels generated between two glass plates and samples electrophoresed vertically. However, polyacrylamide gels cast in trays and electrophoresed horizontally offers several advantages. For example, horizontal gels used to analyze complexes between fluorescent RNA substrates and specific proteins can be imaged multiple times as electrophoresis progresses. This provides the unique opportunity to monitor RNA-protein complexes at several points during the experiment. In addition, horizontal gel electrophoresis makes it possible to analyze many samples in parallel. This can greatly facilitate time course experiments as well as analyzing multiple reactions simultaneously to compare different components and conditions. Here we provide a detailed protocol for generating and using horizontal native gel electrophoresis for analyzing RNA-Protein interactions.

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