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JoVE Journal
Immunology and Infection
Author Produced

This content is Open Access.

Creation of a Dense Transposon Insertion Library Using Bacterial Conjugation in Enterobacterial Strains Such As Escherichia Coli or Shigella flexneri
 

Creation of a Dense Transposon Insertion Library Using Bacterial Conjugation in Enterobacterial Strains Such As Escherichia Coli or Shigella flexneri

Article DOI: 10.3791/56216 11:37 min September 23rd, 2017
September 23rd, 2017

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Summary

Presented here is a simple method for creating a high-density transposon insertion library in Escherichia coli or Shigella flexneri using bacterial conjugation. This protocol allows the creation of a collection of hundreds of thousands of unique mutants in bacteria by the random genomic insertion of a transposon.

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Dense Transposon Insertion Library Bacterial Conjugation Enterobacterial Strains Escherichia Coli Shigella Flexneri Tn10 Transposon Random Genomic Insertion Transfer Of The Plasmid Electroporation Sterile Technique Donor Strain Recipient Strain Centrifuge Nitrocellulose Filter LB Agar Plate Antibiotics Blotting Paper
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