Pulse Field Gel Electrophoresis pp 171-189

Part of the Methods in Molecular Biology book series (MIMB, volume 1301) | Cite as

PFGE for Shiga Toxin-Producing Escherichia coli O157:H7 (STEC O157) and Non-O157 STEC

  • Patricia Jaros
  • Muriel Dufour
  • Brent Gilpin
  • Molly M. Freeman
  • Efrain M. Ribot

Abstract

This chapter describes the procedure of generating pulsed-field gel electrophoresis (PFGE) profiles (DNA fingerprints) of Shiga toxin-producing Escherichia coli O157:H7 (STEC O157) and non-O157 STEC strains within 48 h, based on the standardized laboratory protocol developed by the Centers for Disease Control and Prevention, USA. The protocol describes the preparation of agarose plugs containing STEC O157 and non-O157 STEC cells, the digestion of bacterial DNA in the plugs using restriction endonuclease enzymes, and the electrophoresis conditions to generate the characteristic PFGE profiles of STEC O157 and non-O157 STEC isolates.

Key words

E. coli STEC O157 Non-O157 STEC PFGE Standardized protocol Molecular subtyping DNA fingerprint PulseNet 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Patricia Jaros
    • 1
  • Muriel Dufour
    • 2
  • Brent Gilpin
    • 3
  • Molly M. Freeman
    • 4
  • Efrain M. Ribot
    • 4
  1. 1.Molecular Epidemiology and Public Health Laboratory, Hopkirk Research InstituteMassey UniversityPalmerston NorthNew Zealand
  2. 2.Enteric Reference LaboratoryInstitute of Environmental Science & Research LtdUpper HuttNew Zealand
  3. 3.Water GroupInstitute of Environmental Science & Research LtdChristchurchNew Zealand
  4. 4.Enteric Diseases Laboratory BranchCenters for Disease Control and PreventionAtlantaUSA

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