Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4047

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Selection of Poly(A)+ RNA by Oligo(dT)-Cellulose Chromatography

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 100 words of the full text of this article appear below.


INTRODUCTION

Chromatography on oligo(dT) columns is the preferred method for large-scale purification (>25 µg) of poly(A)+ RNA extracted from mammalian cells. Typically, between 1% and 10% of the RNA applied to the oligo(dT) column is recovered as poly(A)+ RNA. Because the method can be frustratingly slow, it is not recommended for purification of poly(A)+ RNA from multiple samples. For this purpose, batch elution (Selection of Poly(A)+ RNA by Batch Chromatography) is the better choice. IMPORTANT: Prepare all reagents used in this protocol with Diethyl pyrocarbonate (DEPC)-treated H2O.


MATERIALS

recipe 2x Oligo(dT)-cellulose column-loading buffer

Ethanol

Optional, please see Step 5.

recipe NaCl . . . [Full Text of this Article]


METHOD


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