Grant: Research Tools and Instruments - Category 1 (<$150,000)

Understanding how genes influence the form, function and fitness of living organisms is a fundamental goal of evolutionary genetics. Gene expression analysis is therefore vital to research progress, yet this analysis is often constrained by limits to the statistical resolution and power of conventional real-time PCR. This limit frequently masks biologically important variation in gene expression and prevents a comprehensive and accurate exploration of the functional genomic underpinning of adaptively complex traits. This shortcoming is circumvented, however, by new technology that allows for the ultra-precise measurement of gene expression. This innovation, made possible by 'digital droplet' polymerase chain reaction (ddPCR) technology provides orders of magnitude more precision and sensitivity than conventional gene expression technology. This break-through for molecular biology is analogous to the relationship between the number of pixels and the resolution of a digital image, allowing biologists to 'see' what genes are doing like never before. This proposal seizes on the advent of new technology to advance the research programs at the Molecular studies in Biodiversity, Ecology and Environmental Sciences facility (MoBEES). This shared facility, within the Department of Biology at Western University, is a molecular research and training laboratory for field biologists and molecular specialists, and provides training to a large number of students and postdocs (more than 70 high-access users over the past five years). The current request for a ddPCR system will exploit a new frontier in PCR, and provide a vital upgrade to the facility's capacity for assessment of nucleic acid quality and for the quantification of gene expression. We fully expect the up-grade to translate into qualitatively superior diagnoses across multiple research programs. Moreover, by integrating the research tool into our widely accessed facility we will train a new generation of students to apply a revolutionary new technique, and help them to discover anew how genes affect the most biologically important traits.