High Throughput Real-Time PCR System for the Microarray Core

描述（由申请人提供）：本提案的具体目的是在阿肯色大学医学科学学院购买应用生物系统公司7900HT高通量实时PCR系统用于微阵列核心。包括博士在内的7位主要研究者的12个nih资助项目需要该仪器。Barger, Griffin, Kern, Lecka-Czernik, Mock, McGehee, Peterson，以及其他Microarray Core客户，包括BRIN和GCRC赞助的项目。该仪器是合理的，因为：(1)在2006-2007年及以后，注意到的用户和Core将调查一些感兴趣的样品（bbb2138）和基因（bbb455），这将需要高通量能力；(2) 7900HT是唯一可以连续运行实时PCR 384孔反应板过夜并收集和合并数据的市售仪器；(3) 7900HT将是阿肯色州唯一的高通量PCR系统；(4) 7900HT提供了最大的灵活性，允许单个研究人员平衡分析成本和劳动强度，以满足他们的需求；(5)现有仪器7700的使用将在2005年达到最大容量，2007年4月以后将不再提供服务合同；(6)增加另一台应用生物系统公司的仪器将提供实验连续性，并将限制主要用户所需的培训时间；(7)预计7900HT将与7700一样可靠，并且能够承受大量用户，7700自2001年以来经历了不到两周的累计停机时间，因此特别适合集成到核心设施中。PI有资格负责该仪器，因为他与人合作撰写了5篇同行评审的研究文章，利用实时PCR，开发了SYBR绿色检测的优化程序，成本是应用生物系统公司推荐的12%，并亲自设计了170多种人类，大鼠和小鼠基因的检测方法，成功率接近100%。咨询委员会将每年或根据需要召开两次会议，以确保公平和合规使用。该机构已承诺超过50,000美元用于管理和维护7900HT PCR系统。该仪器将有利于公共卫生，因为它是一种有价值的研究工具，可以研究生物途径的调节及其失调的病理后果。具体而言，优先项目是在具有广泛意义的领域：阿尔茨海默病和唐氏综合症、代谢综合征和糖尿病、骨质疏松症和骨骼肌脆弱、维生素缺乏症和癌症。

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.