Molecular Biology

Oligodt Columns

Sigma® has prepared product lists for commonly used protocols from Cold Spring Harbor Protocols.

View the protocol on BioSupplyNet.com.

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

Buy this book, M8265

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 Add to Cart
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
0.05% SDS L4390 Sodium dodecyl sulfate for molecular biology
NaCl (5M), Rnase-free S3014 Sodium chloride for molecular biology
Ethanol E7023 Ethanol
NaOH (10 N) 72068 Sodium hydroxide solution, BioChemika Ultra, for molecular biology
Sodium acetate (3 M, pH 5.2) S7899 Sodium acetate buffer solution
Oligo(dt)-cellulose O3131 Oligo(dT)-Cellulose
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.

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