HIGH THROUGHPUT REAL-TIME PCR SYSTEM FOR THE MICROARRAY CORE

该子项目是利用由NIH/NCRR资助的共享仪器赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是用于资助的，不一定是研究者的机构。描述（由申请人提供）：本提案的具体目的是为阿肯色州大学医学科学部的微阵列核心购买应用生物系统7900 HT高重复率实时PCR系统。该仪器是NIH资助的7个主要研究者的12个项目所必需的，包括Barger、Griffin、克恩、Lecka-Czernik、Mock、麦基希、彼得森博士以及其他微阵列核心客户（包括BRIN）和GCRC赞助的项目。该仪器是合理的，因为：（1）著名用户和核心将在2006-2007年及以后研究许多感兴趣的样本（>2138）和基因（>455），这将需要高通量能力;（2）7900 HT是唯一可以连续运行实时PCR 384-孔反应板过夜，收集和合并数据;（3）7900 HT将是阿肯色州唯一的高通量PCR系统;（4）7900 HT提供了最大的灵活性，允许个人研究者平衡分析成本和劳动密集度，以满足他们的需求;（5）现有仪器7700的使用量将于2005年达到最大限度，2007年4月后将不再提供服务合同;（6）增加另一台应用生物系统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;和（7）预计7900 HT将与7700一样可靠，并能容忍大量用户，7700经历了不到2-自2001年以来，累计停机时间为20周，特别适合集成到核心设施中。PI有资格负责该仪器，因为他已经与人合著了5篇利用实时PCR的同行评审研究文章，开发了SYBR绿色测定的优化程序，其成本是Applied Biosystems推荐的12%，并且亲自设计了170多种人类、大鼠和小鼠基因的测定，成功率接近100%。咨询委员会将每年举行两次会议，或根据需要举行会议，以确保公平和合规的使用。该机构已承诺超过50，000美元用于管理和维护7900 HT PCR系统。该仪器将有益于公共卫生，因为它是一种宝贵的研究工具，可用于调查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、代谢综合症和糖尿病、骨质疏松症和骨骼肌脆弱、维生素缺乏症和癌症。

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.