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Molecular Biology
Plasmid DNA Preparation
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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.
View the protocol on BioSupplyNet.com.
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Selected Protocol from "Molecular Cloning: A Laboratory Manual", Third Edition, Joseph Sambrook, David W. Russell.
Preparation of Plasmid DNA by Alkaline Lysis with SDS: Maxipreparation
Joseph Sambrook
Peter Maccallum Cancer Institute and The University of Melbourne, Australia
David W. Russell
University of Texas Southwestern Medical Center, Dallas
ABSTRACT
Plasmid DNA may be isolated from large-scale (500 ml) bacterial cultures by treatment with alkali and SDS. The resulting DNA preparation may be further purified by column chromatography or centrifugation through CsCl-ethidium bromide gradients.
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Products Available for this Protocol
| Protocol Material Description |
Product # |
Product Name |
Add to Cart |
| Alkaline Lysis Solution I |
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| 50 mM glucose |
G5400 |
D-(+)-Glucose |
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| 25 mM Tris-Cl (pH 8.0) |
T5941 |
Trizma® hydrochloride for molecular biology, Biotechnology Performance Certified |
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| 10 mM EDTA (pH 8.0) |
E7889 |
Ethylenediaminetetraacetic acid disodium salt solution for molecular biology |
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| Alkaline Lysis Solution II |
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| 0.2 N NaOH (freshly diluted from a 10 N stock) |
72068 |
Sodium hydroxide solution, BioChemika Ultra, for molecular biology |
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| 1% (w/v) SDS |
L4390 |
Sodium dodecyl sulfate for molecular biology |
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| Alkaline Lysis Solution III |
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| 5 M potassium acetate, 60.0 ml |
P1190 |
Potassium acetate for molecular biology |
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| Glacial acetic acid, 11.5 ml |
A6283 |
Acetic acid, ReagentPlus™ |
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| H2O, 28.5 ml |
W4502 |
Water, Molecular Biology Reagent |
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| General Reagents |
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| Chloramphenicol (34 mg/ml) |
C1863 |
Chloramphenicol Biotechnology Performance Certified |
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| Ethanol |
E7023 |
Ethanol, 200 proof (absolute), for molecular biology |
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| Isopropanol |
I9516 |
2-Propanol for molecular biology, >99% |
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| STE |
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| 10 mM Tris-Cl (pH 8.0) |
T5941 |
Trizma® hydrochloride for molecular biology, Biotechnology Performance Certified |
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| 0.1 M NaCl |
S3014 |
Sodium chloride |
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| 1 mM EDTA (pH 8.0) |
E7889 |
Ethylenediaminetetraacetic acid disodium salt solution for molecular biology |
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| TE (pH 8.0) |
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| 100 mM Tris-Cl (desired pH) |
T5941 |
Trizma® hydrochloride for molecular biology, Biotechnology Performance Certified |
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| 10 mM EDTA (pH 8.0) |
E7889 |
Ethylenediaminetetraacetic acid disodium salt solution for molecular biology |
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| Enzymes |
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| Lysozyme (10 mg/ml) |
L3790 |
Lysozyme from chicken egg white |
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| Media |
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| LB |
L2897 |
LB Agar, powder |
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| YT |
95768 |
YT Agar, BioChemika, for microbiology |
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| Terrific Broth |
T5574 |
Terrific Broth Liquid Medium |
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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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