Protocol

Working with Bacteriophage P1 and Its Cloning Systems

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

This protocol describes methods for recovery and purification of recombinant clones of bacteriophage P1 or PAC DNAs from bacteria. Because of their large size, these DNAs are sensitive to shearing forces and must be handled carefully. This protocol generally yields P1 DNA that works well as a substrate or template in enzymatic reactions.

MATERIALS

E. coli strain transformed with a nonrecombinant bacteriophage P1 or PAC vector (culture)

E. coli strain transformed with a recombinant bacteriophage P1 or PAC vector (culture)

Alkaline lysis solution I

For preparations of plasmid DNA that are to be subjected to further purification by chromatography (please see . . . [Full Text of this Article]