Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4057

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Separation of RNA According to Size: Electrophoresis of Glyoxylated RNA through Agarose Gels

Joseph Sambrook and David W. Russell

This protocol was adapted from Molecular Cloning, 3rd edition, by Joseph Sambrook and David W. Russell. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2001

The first 15% of the full text of this article appears below.


INTRODUCTION

Separation of RNAs according to size is the first stage in northern blotting and hybridization. The method described in this protocol uses glyoxal to denature the RNA, ethidium bromide to stain it, and agarose gel electrophoresis to separate the resulting glyoxal-RNA-ethidium adducts. IMPORTANT: Prepare all reagents used in this protocol with Diethyl pyrocarbonate (DEPC)-treated H2O.


MATERIALS

recipe 10x BPTE electrophoresis buffer

recipe caution DMSO

Purchase a high grade of DMSO (HPLC grade or better).

recipe Glyoxal

Commercial stock solutions of glyoxal (40% or 6 M) contain various hydrated forms of glyoxal, as well as . . . [Full Text of this Article]


METHOD


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