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Chapters
- 00:04Title
- 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
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
Digital PCRCompetitive IndexHigh-throughput AnalysisFitnessSalmonellaQuantificationMixed PopulationMolecular-based TechniquesGenetic MarkersBacterial ChromosomesGenomic DNADilution SeriesDigital PCR SupermixProbe-based ChemistryNo Template ControlsNegative ControlsPositive ControlsDroplet GeneratorThermal CyclerData Analysis Software
