Growing Lambda‐Derived Vectors

Karen Lech1

1 Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts
Publication Name:  Current Protocols in Molecular Biology
Unit Number:  Unit 1.12
DOI:  10.1002/0471142727.mb0112s13
Online Posting Date:  May, 2001
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Abstract

It is often necessary to grow large quantities of lambda‐derived vectors, so that DNA can be made from them. This unit describes a procedure for making a phage stock by plate lysis, and an gives details on how to make a liquid lysate. Storage of phage lysates is described in a .

     
 
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Table of Contents

  • Basic Protocol 1: Making a Stock of Phage by Plate Lysis
  • Alternate Protocol 1: Making a Liquid Lysate
  • Support Protocol 1: Storing Phage Lysates
  • Reagents and Solutions
  • Commentary
  • Key Reference
     
 
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Materials

Basic Protocol 1: Making a Stock of Phage by Plate Lysis

  Materials
  • Fresh lambda top agar (unit 1.1)
  • Fresh lambda plates (unit 1.1), prewarmed to 37°C
  • Suspension medium (SM; unit 1.11)
  • Chloroform
  • 45° to 50°C water bath
  • Capillary tube or toothpick
  • Beckman JA‐21 rotor or equivalent
NOTE: All materials coming into contact with E. coli must be sterile.

Alternate Protocol 1: Making a Liquid Lysate

  Additional Materials
  • LB medium (unit 1.1)
  • Lambda dilution buffer
  • 10 mM MgCl 2/10 mM CaCl 2
  • NZC medium (unit 1.1)
  • Beckman JA‐20 rotor or equivalent
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Figures

Videos

Literature Cited

Key Reference
   Blattner, F.R., Williams, B.G., Blechl, A.E., Denniston‐Thompson, K., Faber, H.E., Furlong, L.‐A., Grunwald, D.J., Kiefer, D.O., Moore, D.D., Schumm, J.W., Sheldon, E.O., and Smithies, O. 1977. Charon phages:Safer derivatives of bacteriophage lambda for DNA cloning. Science 196:161‐169.
  Origin of this liquid lysate procedure.
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