This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The long-term goal of the INBRE Mayville State University research program is to offer undergraduate students a challenging and rewarding experience directed toward biomedical research. Through BRIN Phase I funding, we have begun to incorporate innovative fluorescence technology, which has been developed by a local high tech business with funding from the NSF and NIH. This technology provides a breakthrough in the generation of high quality fluorescence lifetime information. Applications in allele-specific SNP genotyping and real-time PCR decay will be investigated. Our hypothesis is that the fluorescence lifetime methodology will allow multiplexed PCR and SNP detection that can be simultaneously performed on a greater number of targets with less expense, higher sensitivity and selectivity than is provided by equipment and kits that are currently available commercially. Our research plan has four specific aims: 1) characterize the environmental influence on the fluorescent properties of candidate fluorophores (both in their free form and bound to an oligonucleotide) that are candidates for lifetime-based RT-PCR probes. 2) develop methodology using the lifetime fluorescent probes to monitor single and multiplex RT-PCR analysis. 3) develop methodology using the lifetime fluorescent probes to perform SNP detection. 4) Apply the lifetime probe methodology to RT-PCR and SNP genotyping on real-world samples through collaboration with the INBRE groups at Dickinson State University, Turtle Mountain Community College, and the Biology Core Facility at NDSU.
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