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Digital PCR-based Competitive Index for High-throughput Analysis of Fitness in Salmonella
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JoVE Journal Genética
Digital PCR-based Competitive Index for High-throughput Analysis of Fitness in Salmonella

Digital PCR-based Competitive Index for High-throughput Analysis of Fitness in Salmonella

DOI:
10.3791/59630-v

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07:11 min

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May 13, 2019

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,
1Department of Medical Microbiology and Immunology,Creighton University School of Medicine

Capítulos

  • 00:04Título
  • 00:43Validate the Sensitivity and Specificity of Each Primer-probe Set for Each Genomic Barcode Using Digital PCR
  • 03:09Analyze Digital PCR Data and Calculate Absolute Copy Numbers
  • 04:52Results: Representative Digital PCR Results of Diluted AA-barcoded gDNA
  • 06:28Conclusion

Summary

Tadução automática

Tadução automática

This molecular-based approach for determining bacterial fitness facilitates precise and accurate detection of microorganisms using unique genomic DNA barcodes that are quantified via digital PCR. The protocol describes calculating the competitive index for Salmonella strains; however, the technology is readily adaptable to protocols requiring absolute quantification of any genetically-malleable organism.

Tags

Keywords: Digital PCRCompetitive IndexHigh-throughput AnalysisFitnessSalmonellaQuantificationMixed PopulationMolecular-based TechniquesGenetic MarkersBacterial ChromosomesGenomic DNADilution SeriesDigital PCR SupermixProbe-based ChemistryNo Template ControlsNegative ControlsPositive ControlsDroplet GeneratorThermal CyclerData Analysis Software

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