Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot3901

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Preparation of Plasmid DNA by Alkaline Lysis with SDS: Midipreparation

Joseph Sambrook and David W. Russell

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

The first 15% of the full text of this article appears below.


INTRODUCTION

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


MATERIALS

recipe 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.

recipe Alkaline lysis solution II

recipe Alkaline lysis solution III

Antibiotic . . . [Full Text of this Article]


METHOD


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