Purification of Cosmid and Plasmid Clones

John H. Weis1

1 Harvard Medical School, Boston, Massachusetts
Publication Name:  Current Protocols in Molecular Biology
Unit Number:  Unit 6.6
DOI:  10.1002/0471142727.mb0606s17
Online Posting Date:  May, 2001
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Abstract

Cosmid‐ or plasmid‐bearing colonies that are identified by hybridization are purified by spreading the cosmids or plasmids on an agar plate and repeating the colony hybridization.

     
 
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Table of Contents

  • Commentary
     
 
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Materials

Basic Protocol 1:

  Materials
  • Cold LB medium containing antibiotic (unit 1.1)
  • LB plates containing antibiotic (unit 1.1)
  • Round toothpicks (unit 1.1)
  • Nitrocellulose membrane filters
  • Spreader (unit 1.3)
  • Additional reagents and equipment for plating bacteria (unit 1.3), plating and transferring plasmid/cosmid libraries (unit 6.2), plasmid minipreps (unit 1.6), and autoradiography ( appendix 3A)
NOTE: All materials coming into contact with E. coli must be sterile.
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Figures

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Literature Cited

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