Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot3933

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Transformation of E. coli by Electroporation

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 100 words of the full text of this article appear below.


INTRODUCTION

Electrocompetent bacteria are prepared by growing cultures to mid-log phase, washing the bacteria extensively at low temperature, and then resuspending them in a solution of low ionic strength containing glycerol. DNA is introduced during exposure of the bacteria to a short high-voltage electrical discharge.


MATERIALS

recipe GYT medium, ice cold

recipe Glycerol (10% v/v) (molecular biology grade), ice cold

recipe LB medium, prewarmed to 37°C

Plasmid DNA (recombinant plasmid)

Construct using one of the methods described in Directional Cloning into Plasmid Vectors, Attaching Adaptors to Protruding Termini, Blunt-ended Cloning into Plasmid Vectors, Dephosphorylation of Plasmid DNA, Addition of Synthetic Linkers . . . [Full Text of this Article]


METHOD


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