|
|
Molecular Biology
BAC DNA - Large-scale Cultures
|
|
|
|
|
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.
|
|
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.
back to top
View all available Cold Spring Harbor Laboratory Press 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.
|
|
