Digital PCR infrastructure to enhance research and HQP training in biology

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Quantitative PCR (qPCR) revolutionized many fields within ecology and evolution, molecular biology, agriculture, medicine, and civil engineering, allowing for rapid quantification of nucleic acids in a range of biological samples. Yet quantification of a target template DNA using traditional qPCR relies on a standard dilution series, where accuracy decreases with lower target abundance, standardization among experiments can be challenging, and there can be high rates of false negatives and positives. For many applications in our laboratories requiring detection and quantification of low copy number target DNA template (e.g. detections of rare alleles, early stages of biological invasions, rare species of conservation concern, or ancient eDNA in lake sediments), digital PCR (dPCR) provides an important technological leap forward, allowing for direct counts of individual target templates. In the Department of Biology at Queen's University, we face two challenges: 1) For many projects that need highly-sensitive, quantitative DNA detection, the limitations of traditional qPCR are compromising research progress and HQP training, our ability to undertake novel research, and our ability to publish in the highest-quality journals. 2) Irrespective of the limitations of traditional qPCR, we have only one over-subscribed qPCR machine of any kind for our entire department of 31 full-time faculty, >120 graduate students, and >60 undergraduate thesis students. To rectify these issues, we propose to purchase a high precision BioRad QX200 Droplet Digital PCR workstation that will be housed in our shared departmental molecular core lab, and made available to all within our department, with priority given to co-applicants on this proposal.