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The following Cold Spring Harbor Laboratory Press protocols are brought to you by BioSupplyNet in a partnership with Sigma-Aldrich. A list of products is available to use in each protocol.
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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
ABSTRACT
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 |
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| Chloroform |
C2432 |
Chloroform |
|
| Ethanol |
E7023 |
Ethanol |
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| 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 |
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| Proteinase K |
P2308 |
Proteinase K from Tritirachium album |
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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.
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