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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
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Buy this book, M8265
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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 |
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| Ethanol |
E7023 |
Ethanol |
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| Isopropanol |
I9516 |
2-Propanol for molecular biology |
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| Phenol:chlorofrom (1:1, v/v) |
P2069 |
Phenol:Chloroform:Isoamyl Alcohol 25:24:1 |
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| 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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| 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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DNA markers for pulsed-field gel electrophoresis
| Product Code |
Product Name |
Add to Cart |
| D2291 |
Pulse Marker™ 0.1-200 kb |
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| D2416 |
Pulse Marker™ 50-1,000 kb |
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| D4658 |
Pulse Marker™ 225-2,200 kb |
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Note: As an alternative, the entire protocol could be accomplished with product NA0100.
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