NOVEL FLUORESCENCE METHODS FOR BIOMEDICAL APPLICATIONS

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 INBRE梅维尔州立大学研究计划的长期目标是为本科生提供针对生物医学研究的具有挑战性和有益的经验。 通过BRIN第一阶段的资助，我们已经开始采用创新的荧光技术，该技术是由当地一家高科技企业在NSF和NIH的资助下开发的。 该技术为高质量荧光寿命信息的产生提供了突破。 将研究在等位基因特异性SNP基因分型和实时PCR衰减中的应用。 我们的假设是，荧光寿命的方法将允许多重PCR和SNP检测，可以同时进行更多的目标，更少的费用，更高的灵敏度和选择性比目前市售的设备和试剂盒提供。 我们的研究计划有四个具体目标： 1)表征环境对候选荧光团（游离形式和与寡核苷酸结合的形式）的荧光性质的影响，所述候选荧光团是基于寿命的RT-PCR探针的候选物。 2)开发使用寿命荧光探针的方法，以监测单一和多重RT-PCR分析。 3)开发使用寿命荧光探针进行SNP检测的方法。 4)通过与迪金森州立大学、龟山社区学院和NDSU生物学核心设施的INBRE小组合作，将寿命探针方法应用于真实世界样本的RT-PCR和SNP基因分型。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.