Protocol

Preparation of Plasmid DNA by Alkaline Lysis with SDS: Midipreparation

This protocol was adapted from Molecular Cloning, 3rd edition, by Joseph Sambrook and David W. Russell. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2001

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INTRODUCTION

Plasmid DNA is isolated from intermediate-scale (20-50 ml) bacterial cultures by treatment with alkali and SDS.

MATERIALS

Alkaline lysis solution I

For preparations of plasmid DNA that are to be subjected to further purification by chromatography (please see Purification of Plasmid DNA by Chromatography), sterile Alkaline lysis solution I may be supplemented just before use with the appropriate volume of 20 mg/ml DNase-free RNase A (pancreatic RNase) to give a final concentration of 100 µg/ml.

Alkaline lysis solution II

Alkaline lysis solution III

Antibiotic . . . [Full Text of this Article]

METHOD