PERKIN ELMER 7700 SEQUENCE DETECTOR

PERKIN ELMER 7700 序列检测器

• 批准号: 6291660
• 负责人: JOHN A OLSCHOWKA
• 金额: $ 9.53万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-15 至 2002-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6291660
• 关键词: biomedical equipment purchase; polymerase chain reaction

The objective of this proposal is to seek support for the purchase of a real-time PCR machine (PE 7700 Sequence Detector). The polymerase chain reaction (PCR) has revolutionized the detection of DNA and RNA. As little as a single copy of a particular sequence can be specifically amplified and detected. Theoretically, there is a quantitative relationship between amount of starting target sequence and amount of PCR product at any given cycle. In practice, though, it is a common experience for replicate reactions to yield different amounts of PCR product. The development of real-time quantitative PCR has eliminated the variability traditionally associated with quantitative PCR, thus allowing the routine and reliable quantification of PCR products. J.A. Olschowka (principal investigator) and M.K. O'Banion (major user) routinely use competitive quantitative RT-PCR in their NIH-funded work examining inflammation in the central nervous system. Other major users, all of whom are funded by NIH, propose to use the PE 7700 for quantifying DNA and/or RNA in a variety of settings. D.A. Figlewicz will use the instrument in expression and mapping studies of candidate genes potentially involved in Facioscapulohumeral Dystrophy. J. Finkelstein studies cytokine and chemokine expression in acute lung injury and progressive pulmonary fibrosis. Currently using RNAse protection assays (RPA) to analyze mRNA levels, his work will be accelerated by real-time PCR. Dr. Rodier will use the instrument in her molecular studies of autism. The sensitivity of the instrument will be especially important to her when working with small amounts of mouse embryo tissue. Dr. Phipps studies the role of CD 40 in lung injury as well as the regulation of immunity by prostaglandins. Again, analysis of mRNA levels in these studies will be enhanced over current RPA methods in terms of speed, reliability and reproducibility. The instrument will be housed in the Molecular Biology core facility within the Department of Neurobiology and Anatomy. This core facility is the responsibility of the J. A. Olschowka (the Principal Investigator) who will oversee the daily use of the instrument. A committee made up of the Major Users will provide long-term guidance about the use of the instrument. The institution will provide a room of approximately 350 sq feet to house the instrument, ancillary equipment required for PCR, and cost-sharing of service contracts.

该提案的目的是寻求购买实时PCR机（PE 7700序列检测器）的支持。聚合酶链反应（PCR）已彻底改变了DNA和RNA的检测。只能特异性放大和检测到特定序列的单个副本。从理论上讲，在任何给定循环中的起始目标序列和PCR产物的量之间存在定量关系。但是，在实践中，复制反应的常见经验是产生不同量的PCR产品。实时定量PCR的开发消除了传统上与定量PCR相关的可变性，从而允许对PCR产物的常规和可靠的量化。 J.A. Olschowka（首席研究员）和M.K. O'Banion（主要用户）通常在其NIH资助的工作中使用有竞争力的定量RT-PCR检查中枢神经系统的炎症。其他主要用户（所有人都由NIH资助）建议在各种环境中使用PE 7700来量化DNA和/或RNA。 D.A. Figlewicz将使用该仪器在表达和映射研究中可能涉及facioscapulohumeral营养不良的候选基因。 J. Finkelstein研究急性肺损伤和进行性肺纤维化的细胞因子和趋化因子的表达。目前使用RNase保护分析（RPA）来分析mRNA水平，他的工作将通过实时PCR加速。罗德尔博士将在其自闭症分子研究中使用该仪器。使用少量小鼠胚胎组织时，仪器的敏感性对她尤为重要。 Phipps博士研究CD 40在肺损伤中的作用以及前列腺素对免疫的调节。同样，就速度，可靠性和可重复性而言，对当前RPA方法的mRNA水平的分析将增强。该仪器将安置在神经生物学和解剖学系的分子生物学核心设施中。该核心设施是J. A. Olschowka（主要研究人员）的责任，他们将监督该工具的日常使用。由主要用户组成的委员会将提供有关该工具使用的长期指导。该机构将提供约350平方英尺的房间，以容纳乐器，PCR所需的辅助设备以及服务合同的成本分配。