Transformation of E. coli by Electroporation

Sherie L. Morrison1

1 University of California Los Angeles, Los Angeles, California
Publication Name:  Current Protocols in Immunology
Unit Number:  Appendix 3N
DOI:  10.1002/0471142735.ima03ns21
Online Posting Date:  May, 2001
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Abstract

Using electroporation to transform Escherichia coli results in transformation efficiencies greater than can be obtained using the best chemical methods. It is easy to obtain transformation efficiencies 108 per milligram DNA and efficiencies of 1010 have been reported. This appendix describes a procedure for electroporation that can be used to transform many different types of bacteria.

     
 
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Table of Contents

  • Reagents and Solutions
  • Commentary
  • Literature Cited
     
 
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Materials

Basic Protocol 1:

  Materials
  • recipeLB medium (unit 10.18)
  • Transformation competent E. coli (e.g., HB101 or XL1 blue, Stratagene)
  • recipeSterile H 2O (filter sterilize, see recipe), 4°C
  • 15% (v/v) glycerol, sterile, 4°C
  • 5 pg to 0.5 µg/µl DNA for transformation
  • recipeSOC medium (see recipe)
  • 100 mg/ml isopropyl β‐D‐thiogalactopyranoside (IPTG) in H 2O (optional)
  • 15 mg/ml 5‐bromo‐4‐chloro‐3‐indoyl‐β‐D‐galactopyranoside (Xgal) in dimethylformamide (optional)
  • recipeLB plates with appropriate antibiotic (see recipe)
  • 50‐ml conical tube, sterile
  • Shaking incubator, 37°C
  • Fernbach or 2‐liter flask
  • 250‐ml centrifuge bottles
  • Sorval or similar centrifuge with GSA rotor to accommodate 250‐ml bottles, 4°C
  • 5‐ml pipets
  • Microcentrifuge, 4°C
  • Dry ice/ethanol bath
  • Electroporation cuvette, sterile
  • Electroporation apparatus
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Figures

Videos

Literature Cited

Literature Cited
   Dower, W.J., Miller, J.F., and Ragsdale, C.W. 1988. High efficiency transformation of E. coli by high voltage electroporation. Nucl. Acids Res. 16:6127‐6145.
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