7700 SEQUENCE DETECTOR AND BIOMEK WORKSTATION

7700 序列检测器和 BIOMEK 工作站

• 批准号: 2803494
• 负责人: PETER J DAVIES
• 金额: $ 17.38万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-15 至 2000-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2803494
• 关键词: biomedical equipment; biomedical equipment purchase; nucleic acid quantitation /detection; nucleic acid sequence; polymerase chain reaction; robotics

A major limitation in current investigations of the cellular and molecular basis of hormone and drug action is the lack of convenient techniques for the precise and reliable quantitation of specific gene transcripts in biological systems. The current techniques available for the measurement of specific mRNA's are either very insensitive and imprecise (Norther blots_ or require multiple complex experimental manipulations (RNAase protection or competitive Q-PCR). "Real-time" Quantitative RT-PCR offers a novel solution to this problem by facilitating the direct measurement of the progress of a PCR amplification in "real time". By gathering data during the exponential phase of the PCR amplification and by eliminating the need for electrophoresis and quantitation of individual amplification reactions, this system allows for the precise, reproducible and simple quantitation of specific mRNAs. ABI has recently developed a multiplexed instrument capable of supporting "real-time" PCR reactions in a 96-well array format, allowing for the high throughput analysis of multiple transcripts. When coupled with a Beckman robotic workstation, this instrument can be used to generate quantitative data on large numbers of transcripts in a single day. We propose to establish a core Q-PCR facility comprised of these instruments under the supervision of a skilled research scientists that will be available to participating investigators for the measurement of specific transcripts of interest to particular research projects. The availability of such a facility will not only extend the scope of current funded research projects but will provide the opportunity to develop novel experimental approaches not previously feasible because of the logistical limitations of conventional assay procedures.

目前对激素和药物作用的细胞和分子基础研究的主要局限性是缺乏对生物系统中特定基因转录物进行精确和可靠定量的方便技术。当前可用于测量特定mRNA的技术要么非常不敏感和不精确（Norther blots_或需要进行多种复杂的实验操作（RNAase保护或竞争性Q-PCR）。“实时”定量RT-PCR可以通过促进PCR逐步启动“实时数据”的直接测量来促进“实时测量”。消除对电泳的需求和对单个扩增反应的定量，该系统允许对特定mRNA的精确，可重复的和简单的定量。在一天中的大量成绩单上。我们建议在熟练的研究科学家的监督下建立由这些工具组成的核心Q-PCR设施，该工具将可以向参与研究人员提供，以测量特定研究项目的特定成绩单。这种设施的可用性不仅会扩大当前资助的研究项目的范围，而且还将为开发新型实验方法提供的机会，因为传统测定程序的后勤局限性，以前不可行。