Attention:

Certain features of Sigma-Aldrich.com will be down for maintenance the evening of Friday August 18th starting at 8:00 pm CDT until Saturday August 19th at 12:01 pm CDT.   Please note that you still have telephone and email access to our local offices. We apologize for any inconvenience.

Molecular Biology

Plasmid DNA Preparation

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.

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.

 

Buy this book, M8265

Products Available for this Protocol

Protocol Material Description Product #  Product Name Add to Cart
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
General Reagents      
Chloramphenicol (34 mg/ml) C1863 Chloramphenicol Biotechnology Performance Certified
Ethanol E7023 Ethanol, 200 proof (absolute), for molecular biology
Isopropanol I9516 2-Propanol for molecular biology, >99%
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
TE (pH 8.0)      
100 mM Tris-Cl (desired pH) T5941 Trizma® hydrochloride for molecular biology, Biotechnology Performance Certified
10 mM EDTA (pH 8.0) E7889 Ethylenediaminetetraacetic acid disodium salt solution for molecular biology
Enzymes      
Lysozyme (10 mg/ml) L3790 Lysozyme from chicken egg white
Media      
LB L2897 LB Agar, powder
YT 95768 YT Agar, BioChemika, for microbiology
Terrific Broth T5574 Terrific Broth Liquid Medium

back to top
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.