Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4048

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Selection of Poly(A)+ RNA by Batch 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 15% of the full text of this article appears below.


INTRODUCTION

When many RNA samples are to be processed or when working with small amounts (<50 µg) of total mammalian RNA, the technique of choice is batch chromatography on oligo(dT)-cellulose. The method described in this protocol uses a combination of temperature and ionic strength to maximize binding and recovery of polyadenylated RNA. IMPORTANT: Prepare all reagents used in this protocol with Diethyl pyrocarbonate (DEPC)-treated H2O.


MATERIALS

recipe caution Ammonium acetate (10 M)

Ice-cold water

recipe RNA absorption/washing buffer

This buffer is TES containing 0.5 M . . . [Full Text of this Article]


METHOD


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