Molecular Biology

Purification of PCR Products

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

Purification of PCR Products in Preparation for Cloning

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

The residual enzymatic activity of thermostable DNA polymerases that survive the rigors of PCR can compromise subsequent enzymatic reactions. This protocol describes how to use proteinase K to destroy thermostable enzymes and to purify amplified DNA in preparation for cloning.

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Products Available for this Protocol

Protocol Material Description Product #  Product Name Add to Cart
Ammonium acetate (10M) A1542 Ammonium acetate
Chloroform C2432 Chloroform
Ethanol E7023 Ethanol
Phenol:chlorofrom (1:1, v/v) P2069 Phenol:Chloroform:Isoamyl Alcohol 25:24:1 Saturated with 10 mM Tris, pH 8.0, 1 mM EDTA
TE (pH 8.0) T9285 Tris-EDTA Buffer 100× Concentrate
Proteinase K P2308 Proteinase K from Tritirachium album

Note 1: For Step 6 of the protocol, you may use the GenElute™ PCR Clean-Up Kit, NA1020.

Note 2: As an alternative, the entire protocol could be accomplished with procuct NA1020, GenElute™ PCR Clean-Up Kit.

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Product Association Disclaimer: The Sigma-Aldrich products listed for this specific protocol were selected either to match or to supplement the products listed within the actual protocol. The products/reagents from Sigma-Aldrich 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.