Protocol

Preparation of Single-stranded 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

Bacteriophage M13 single-stranded DNA is prepared from virus particles secreted by infected cells into the surrounding medium. The filamentous particles are concentrated by precipitation from a high-ionic-strength buffer with polyethylene glycol. Subsequent extraction with phenol releases the single-stranded DNA, which is then collected by precipitation with ethanol. This protocol is generally used to prepare single-stranded DNA from a small number of M13 isolates. Typically, the yield of single-stranded DNA is 5-10 µg/ml infected culture.

MATERIALS

E. coli cultures infected with bacteriophage M13

Prepare an infected culture as described in Growing Bacteriophage M13 in Liquid Culture. These cultures should be infected . . . [Full Text of this Article]