Amplification of Cosmid and Plasmid Libraries

John H. Weis1

1 Harvard Medical School, Boston, Massachusetts
Publication Name:  Current Protocols in Molecular Biology
Unit Number:  Unit 5.11
DOI:  10.1002/0471142727.mb0511s34
Online Posting Date:  May, 2001
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Abstract

The purpose of amplification is to provide a reagent library that can be used many times. In this protocol, cacteria containing the recombinant clones are grown on agar plates, washed off the plates, and stored in glycerol. An amplified cosmid or plasmid library will contain one library's equivalence in 10 to 100 µl. By producing 100 µl of such a library frozen in 100 1‐ml tubes, over 1000 platings can be expected.

     
 
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Table of Contents

  • Commentary
     
 
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Materials

Basic Protocol 1:

  Materials
  • LB plates containing appropriate antibiotic (unit 1.1)
  • LB medium (unit 1.1)
  • Sterile glycerol
  • Nitrocellulose membrane filters (Millipore HATF)
  • Sterile rubber policeman
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Figures

Videos

Literature Cited

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