Oligodt Columns

Product lists for commonly used protocols from Cold Spring Harbor Protocols

Selected Protocol from "Molecular Cloning: A Laboratory Manual", Third Edition, Joseph Sambrook, David W. Russell.

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

Joseph Sambrook -- Peter Maccallum Cancer Institute and The University of Melbourne, Australia
David W. Russell -- University of Texas Southwestern Medical Center, Dallas

ABSTRACT
Chromatography on oligo(dT) columns is the preferred method for large-scale purification (>25 mg) of poly(A)⁺ RNAextracted 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 (Protocol 7.4) is the better choice.

IMPORTANT: Prepare all reagents used in this protocol with DEPC-treated H₂O.

Products Available for this Protocol

Protocol Material Description	Product #	Product Name
2x Column-loading buffer
40 mM Tris-HCl (pH 7.6)	T5941	Trizma® hydrochloride for molecular biology, Biotechnology Performance Certified
1 mM NaCl	S3014	Sodium chloride for molecular biology
2 mM EDTA (pH 8.0)	E5134	Ethylenediaminetetraacetic acid disodium salt solution for molecular biology
Elution Buffer
10 mM Tris-HCl (pH 7.6)	T5941	Trizma® hydrochloride for molecular biology, Biotechnology Performance Certified
1 mM EDTA (pH 8.0)	E5134	Ethylenediaminetetraacetic acid disodium salt solution for molecular biology
NaCl (5M), Rnase-free	S3014	Sodium chloride for molecular biology
Ethanol	E7023	Ethanol
Sodium acetate (3 M, pH 5.2)	S7899	Sodium acetate buffer solution
pH Paper (pH Test Strips)	P4786	pH Test strips

Note: As an alternative, the entire protocol could be accomplished with product MRN10 or MRN70, the GenElute™ mRNA Miniprep Kits.

Product Association Disclaimer: Products listed for this specific protocol were selected either to match or to supplement the products listed within the actual protocol. Our products and reagents have been qualified for usage, but may not have been validated for this specific application. Please refer to the detailed product description on the usage of specific products of interest.