HIGH THROUGHPUT REAL-TIME PCR SYSTEM FOR THE MICROARRAY CORE

这个子项目是利用由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。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是用于拨款的，而不一定是用于调查人员的机构。描述(由申请人提供)：本提案的具体目标是为阿肯色州医学大学的微阵列核心购买一台应用生物系统7900HT高通量实时聚合酶链式反应系统。该仪器是NIH资助的7个主要研究人员的12个项目所必需的，这些项目包括Barger博士、Griffin博士、Kern博士、Lecka-Czernik博士、Mock博士、McGehee博士、Peterson博士和包括Brin在内的其他微阵列核心客户以及GCRC赞助的项目。该仪器是合理的，因为：(1)所提到的用户和核心将在2006-2007年调查一些样本(&gt；2138)和感兴趣的基因(&gt；455)，这将需要高通量能力；(2)7900HT是唯一可以连夜连续运行实时PCR384井反应板并收集和合并数据的商业仪器；(3)7900HT将是阿肯色州唯一的高通量聚合酶链式反应系统；(4)7900HT提供了最大的灵活性，允许个人调查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月之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；以及(7)预计7900HT将与自2001年以来累计停机时间不到两周的7700一样，对大量用户具有同样的可靠性和容忍度，使其特别适合整合到核心设施中。PI有资格负责该仪器，因为他利用实时PCR与人合著了5篇同行评议的研究文章，开发了SYBR绿色分析的优化程序，其成本是应用生物系统公司推荐的程序的12%，并亲自设计了170多个人类、大鼠和小鼠基因的分析，成功率接近100%。咨询委员会将每年举行两次会议，或视需要举行会议，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7900HTPCR系统的管理和维护。该仪器将有益于公众健康，因为它是一种宝贵的研究工具，用于调查生物途径的调节及其失调的病理后果。具体而言，优先项目涉及具有广泛意义的领域：阿尔茨海默氏症和唐氏综合症、代谢综合征和糖尿病、骨质疏松症和骨骼肌虚弱、维生素缺乏和癌症。

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.