Protocol

Preparation of Double-stranded (Replicative Form) Bacteriophage M13 DNA

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

The double-stranded replicative form (RF) of bacteriophage M13 is isolated from infected cells using methods similar to those used to purify plasmid DNA. Several micrograms of RF DNA can be isolated from a 1-2-ml culture of infected cells.

MATERIALS

E. coli culture infected with bacteriophage M13

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 . . . [Full Text of this Article]