Validate the Sensitivity and Specificity of Each Primer-probe Set for Each Genomic Barcode Using Digital PCR
3:09
Analyze Digital PCR Data and Calculate Absolute Copy Numbers
4:52
Results: Representative Digital PCR Results of Diluted AA-barcoded gDNA
6:28
Conclusion
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This technique enables accurate and precise quantification of microorganisms from a mixed population of highly similar strains using rapid and high throughput molecular-based techniques. Ultimately, in enabling comparisons of fitness to be made be
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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.