HIGH THROUGHPUT REAL-TIME PCR SYSTEM FOR THE MICROARRAY CORE

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This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation 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 grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): The specific aim of this proposal is the purchase of an Applied Biosystems 7900HT High Throughput Real-Time PCR System for the Microarray Core at the University of Arkansas for Medical Sciences. The instrument is required for 12 NIH-funded projects of 7 principle investigators including Drs. Barger, Griffin, Kern, Lecka-Czernik, Mock, McGehee, Peterson, and other Microarray Core clientele including BRIN, and GCRC sponsored projects. The instrument is justified because: (1) The noted users and Core will investigate a number of samples (>2138) and genes (>455) of interest in 2006-2007 and beyond that will necessitate high-throughput capabilities; (2) the 7900HT is the only commercially available instrument that can continuously run real-time PCR 384-Well reaction plates overnight and collect and merge the data; (3) the 7900HT will be the only high-throughput PCR system in the state of Arkansas; (4) the 7900HT offers the greatest flexibility to allow individual investigators to balance assay cost and labor intensiveness to meet their needs; (5) use of the current instrument, the 7700, will reach maximum capacity in 2005 and service contracts will not be offered after April 2007; (6) adding another Applied Biosystems instrument will provide experimental continuity and will limit the training time needed for major users; and (7) it is expected that the 7900HT will be just as reliable for and tolerant of a large number of users as the 7700 which experienced less than 2-weeks cumulative down time since 2001 making it especially appropriate for integration into a core facility. The PI is qualified to be responsible for the instrument as he has co-authored 5 peer-reviewed research articles utilizing real-time PCR, developed optimization procedures for SYBR green assays that cost 12% of those recommended by Applied Biosystems and has personally designed assays for more than 170 human, rat, and mouse genes with nearly a 100% success rate. An Advisory Committee will meet twice yearly or as needed to assure equitable and compliant usage. The institution has committed more than $50,000 to the management and maintenance of the 7900HT PCR System. The instrument will benefit public health as it is a valuable research tool to investigate regulation of biological pathways and the pathological consequences of their dysregulation. Specifically, priority projects are in areas of widespread significance: Alzheimer's disease and Down's syndrome, metabolic syndrome and diabetes, osteoporosis and skeletal muscle frailty, vitamin deficiency, and cancer.