Listeria monocytogenes: Cultivation and Laboratory Maintenance

Grant S. Jones1, Sarah E.F. D'Orazio1

1 University of Kentucky, Lexington, Kentucky
Publication Name:  Current Protocols in Microbiology
Unit Number:  Unit 9B.2
DOI:  10.1002/9780471729259.mc09b02s31
Online Posting Date:  November, 2013
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This unit describes general procedures for the lab cultivation and storage of the Gram‐positive facultative intracellular bacterium Listeria monocytogenes. The basic protocols are relevant for a wide scope of applications including microbial genetics and both in vitro and in vivo infection studies. Commonly used L. monocytogenes strains, serotypes, and growth parameters are also discussed. Curr. Protoc. Microbiol. 31:9B.2.1‐9B.2.7. ©2013 by John Wiley & Sons, Inc.

Keywords: Listeria monocytogenes; Gram‐positive; intracellular pathogen; listeriosis

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Table of Contents

  • Introduction
  • Basic Protocol 1: Growth of L. monocytogenes in Liquid Medium
  • Basic Protocol 2: Short‐Term Storage of Listeria Aliquots
  • Basic Protocol 3: Long‐Term Storage of L. monocytogenes
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
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Basic Protocol 1: Growth of L. monocytogenes in Liquid Medium

  • L. monocytogenes (from frozen stock, agar stab, or freeze‐dried pellet)
  • Sterile BHI agar (Difco) plates
  • Sterile BHI broth (Difco)
  • Incubator (with orbital shaker if desired) set to desired temperature (see Commentary)

Basic Protocol 2: Short‐Term Storage of Listeria Aliquots

  • Sterile BHI broth (Difco)
  • L. monocytogenes grown on BHI agar
  • Sterile BHI agar (Difco) plates
  • Incubator (with orbital shaker if desired) set to desired temperature (see Commentary)

Basic Protocol 3: Long‐Term Storage of L. monocytogenes

  • L. monocytogenes grown on BHI agar or in BHI broth culture
  • 40% or 20% glycerol (v/v) BHI broth, sterile (see reciperecipes)
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Literature Cited

Literature Cited
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