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.

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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

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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