There is a tremendously useful-actually, essential-instrument and technology for molecular biologists that should be made available at the University of Minnesota. Toward this goal, this grant provides the integrated instrumentation system to establish a """"""""Real-time Quantitative PCR Core Facility."""""""" With a high level of sensitivity, the presence of any gene can be identified and quantitated, as well as it expression by reverse transcriptase-PCR (RT-PCR). As initially demonstrated by preliminary studies related to a gene therapy clinical trial, this instrumentation fills a very significant scientific need to accurately quantitate the presence of a specific gene, and to measure its expression, in very sensitive arrays. However, the technology and automated natured of the system will accommodate a high throughput capability, with methods that are readily generalizable to assess virtually any gene, in any biologic system. The demand for these capabilities was quickly recognized by 37 investigators who were enlisted as the Major User group at this institution, and whose projects are outlined in this application. The technical expertise to disseminate this methodology will be provided by the PI and other three experienced Ph.D.-level molecular biologist, who having developed assays for their own projects, will provide consultation to other investigators as well as provide day-to-day supervision of the instrumentation. The management plan for the instrument will be administered by the PI and an Advisory Committee of experienced scientists whose cumulative experience includes the administration of other core facilities, GMP manufacturing of gene therapy vectors, and CLIA-certified/CAP-certified clinical testing. This plan includes: (1) provision of expert monitoring and maintenance of the instrumentation; (2) evaluation and assurance of a fair distribution and instrument time by many users; (3) a means to offset the expense of maintenance contracts and other incidental costs by a small charge to the users; and (4) on-line scheduling with electronic archiving of process documentation and data. A strong institutional commitment to establish and maintain this core facility has been articulated by the institution's Vice President for Health Sciences, Dean of the Medical School and Department Head; they have indicated that the core facility will be supported and is directly in line with the mission to advance the University to the top of academic institutions by giving molecular biology research the highest priority. The overall benefit of developing a """"""""Real-time Quantitative PCR Core Facility"""""""" extends beyond the 37 investigators who have made a commitment as Major Users; without a doubt, every molecular biologist at this institution has an important application for the system. The automated system of robotic 96-well tray reaction set-up, married to the real-time fluorometric Taqman analytical component, provides a high throughput integrated system. This core facility will serve as a model to the broader academic community demonstrating how academicians will be able to exploit the important technological advancement of quantitative PCR.
