Plating Lambda Phage to Generate Plaques

Karen Lech1, Roger Brent1

1 Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts
Publication Name:  Current Protocols in Molecular Biology
Unit Number:  Unit 1.11
DOI:  10.1002/0471142727.mb0111s13
Online Posting Date:  May, 2001
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Abstract

This unit provides detailed protocols for isolating a single plaque by titering serial dilutions or streaking on a lawn of cells. A discussion of phage transfection and in vitro packaging is also included.

     
 
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Table of Contents

  • Basic Protocol 1: Isolating a Single Plaque by Titering Serial Dilutions
  • Commentary
  • Basic Protocol 2: Isolating Single Plaques by Streaking on a Lawn of Cells
  • Basic Protocol 3: Phage Transfection and In Vitro Packaging
  • Reagents and Solutions
  • Literature Cited
     
 
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Materials

Basic Protocol 1: Isolating a Single Plaque by Titering Serial Dilutions

  Materials
  • Lambda broth (unit 1.1)
  • 0.2% maltose
  • 10 mM MgSO 4
  • Lambda top agar (unit 1.1)
  • recipeSuspension medium (SM)
  • Fresh lambda plates (unit 1.1), prewarmed to 37°C
  • Microwave oven or boiling water bath
  • 45° to 50°C water bath
  • 8 × 80–mm tubes
  • 37°C water bath or heat block
  • Capillary tubes or toothpicksNOTE: All materials coming into contact with E. coli must be sterile.

Basic Protocol 2: Isolating Single Plaques by Streaking on a Lawn of Cells

  Materials
  • LB medium (unit 1.1)
  • Lambda top agar (unit 1.1)
  • Rich plate (unit 1.1), prewarmed to 37°C
  • 32‐G platinum wire loop or sterile 1 1/2 × 1/8 in. strips of paper
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Figures

Videos

Literature Cited

Literature Cited
   Enquist, L. and Sternberg, N. 1979. Packaging of bacteriophage λ in vitro. Meth. Enzymol. 68:281‐298.
   Stent, G.S. 1971. Molecular Genetics: An Introductory Narrative, W.H. Freeman, NY.
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