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Molecular Biology

BAC DNA - Large-scale Cultures

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.

Isolation of BAC DNA from Large-scale Cultures

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
The procedure for isolation of BAC DNA is scaled up to accommodate 500-ml cultures, which, on average, yield 20-25 µg of purified BACDNA.

Products Available for this Protocol

Protocol Material Description Product #  Product Name Add to Cart
Reagents      
Ethanol E7023 Ethanol
Isopropanol I9516 2-Propanol for molecular biology
Phenol:chlorofrom (1:1, v/v) P2069 Phenol:Chloroform:Isoamyl Alcohol 25:24:1
Alkaline Lysis Solution I      
50 mM glucose G5400 D-(+)-Glucose
25 mM Tris-Cl (pH 8.0) T5941 Trizma® hydrochloride for molecular biology, Biotechnology Performance Certified
10 mM EDTA (pH 8.0) E7889 Ethylenediaminetetraacetic acid disodium salt solution for molecular biology
Alkaline Lysis Solution II      
0.2 N NaOH (freshly diluted from a 10 N stock) 72068 Sodium hydroxide solution, BioChemika Ultra, for molecular biology
1% (w/v) SDS L4390 Sodium dodecyl sulfate for molecular biology
Alkaline Lysis Solution III      
5 M potassium acetate, 60.0 ml P1190 Potassium acetate for molecular biology
Glacial acetic acid, 11.5 ml A6283 Acetic acid, ReagentPlus™
H2O, 28.5 ml W4502 Water, Molecular Biology Reagent
STE      
10 mM Tris-Cl (pH 8.0) T5941 Trizma® hydrochloride for molecular biology, Biotechnology Performance Certified
0.1 M NaCl S3014 Sodium chloride
1 mM EDTA (pH 8.0) E7889 Ethylenediaminetetraacetic acid disodium salt solution for molecular biology

DNA markers for pulsed-field gel electrophoresis

Product Code Product Name Add to Cart
D2291 Pulse Marker™ 0.1-200 kb
D2416 Pulse Marker™ 50-1,000 kb
D4658 Pulse Marker™ 225-2,200 kb

Note: As an alternative, the entire protocol could be accomplished with product NA0100.

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View all available product lists for Cold Spring Harbor Laboratory Protocols

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.