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Generering Transposon Indsættelse Biblioteker i Gram-negative bakterier til høj-throughput sekventering
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Biologia
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JoVE Journal Biologia
Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing

Generering Transposon Indsættelse Biblioteker i Gram-negative bakterier til høj-throughput sekventering

DOI:
10.3791/61612-v

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08:19 min

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July 07, 2020

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, ,
1Department of Biology,University of Texas at Arlington

Capitoli

  • 00:04Introduction
  • 00:57Bacterial Mating
  • 02:47Generation of Final Bacterial Mutant Library
  • 04:11DNA Shearing and Poly-C Tail Addition
  • 06:23Results: Transposon Mutant Library
  • 07:31Conclusion

Summary

Traduzione automatica

Traduzione automatica

Vi beskriver en metode til at generere mætte transposon mutant biblioteker i Gram-negative bakterier og efterfølgende forberedelse af DNA amplicon biblioteker for høj-throughput sekventering. Som et eksempel, vi fokuserer på ESKAPE patogen, Acinetobacter baumannii, men denne protokol er modtagelig for en bred vifte af Gram-negative organismer.

Tags

Keywords: Transposon Insertion LibraryGram-negative BacteriaHigh-throughput SequencingEssential GenesAntibiotic ResistanceIn Vivo FitnessMechanical ShearingPoly-CTLRestriction EnzymesAdapter LigationBacterial GeneticsGenotype-phenotype RelationshipsMatingDiaminopimelic AcidGlycerolFrozen Stock

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