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的研究的一个主要限制是缺乏对生物系统中特定基因转录本进行精确和可靠定量的便捷技术。目前可用于测量特定 mRNA 的技术要么非常不灵敏且不精确（Norther blots），要么需要多种复杂的实验操作（RNAase 保护或竞争性 Q-PCR）。“实时”定量 RT-PCR 为此提供了一种新颖的解决方案该系统通过在 PCR 扩增的指数阶段收集数据并消除对各个扩增反应的电泳和定量的需要，促进“实时”直接测量 PCR 扩增的进度，从而解决了这个问题。 ABI 最近开发了一种多重仪器，能够支持 96 孔阵列形式的“实时”PCR 反应，在耦合时可以对多个转录本进行高通量分析。借助贝克曼机器人工作站，该仪器可用于在一天内生成大量转录本的定量数据。我们建议在熟练的研究科学家的监督下建立一个由这些仪器组成的核心 Q-PCR 设施，参与研究人员可以使用该设施来测量特定研究项目感兴趣的特定转录本。这种设施的可用性不仅将扩大当前资助的研究项目的范围，而且将提供开发新的实验方法的机会，而这些方法以前由于传统分析程序的后勤限制而无法实现。