PERKIN ELMER ABI PRISM 7700 SEQUENCE DETECTION SYSTEM

PERKIN ELMER ABI PRISM 7700 序列检测系统

• 批准号: 6054036
• 负责人: Martin Frederick KAGNOFF
• 金额: $ 10.27万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6054036
• 关键词: biomedical equipment purchase; computer assisted sequence analysis; nucleic acid sequence; polymerase chain reaction

This shared instrument grant application requests an ABI PRISM 7700 Sequence Detection System for use by six groups of investigators working in the disciplines of cell signaling, molecular biology, neurobiology, genetics, immunology and microbial pathogenesis for their studies of regulated gene expression and single nucleotide polymorphisms. The system integrates a polymerase chain reaction (PCR)-based thermal cycler with hardware instrumentation employing laser detection of fluorochromes and software. It will provide this group of investigators with a system for real-time quantitation of nucleic acid sequences with accuracy and reproducibility over a wide dynamic range and with high-throughput. By combining thermal cycling, fluorescence detection, and software in a single instrument, the system enables cycle-by-cycle detection of increases in nucleic acid product. In addition, data output is available immediately following sequence amplification without the need for additional purification steps or analysis by gel electrophoresis. Real time quantitative detection of specific nucleic acid sequences is based on the use of fluorescent probes incorporating reporter and quencher dyes on the same probe, with probe cleavage based on the 5' nuclease activity of Taq DNA polymerase. Probe cleavage separates the reporter dye from the quencher, thereby restoring fluorescence. The instrument's spectrograph and a charge-coupled device camera detect fluorescence emission. The system uses a 96-sample well platform, an argon laser to induce fluorescence, and a multiplexed array of optical fibers. The components include: (1) a 7700 sequence detector with built-in thermal cycler with 96-well positions; (2) an apple power Macintosh system with 16Mb RAM, 500-Mb hard disk drive, 120-MHz Power Macintosh, 1.4-Apple Super Drive floppy disk drive, CD ROM drive, 15" color monitor, and Hewlett Packard 1600C color deskjet printer. Sequence detection software used for instrument control, data collection, and automatic data analysis. The ABI PRISM 7700 System will be a crucial core resource for PHS-supported research on the Medical School campus of the Univ. of California at San Diego.

这项共享仪器拨款申请申请了ABI PRISM 7700序列检测系统，供细胞信号、分子生物学、神经生物学、遗传学、免疫学和微生物发病机制等学科的六个研究小组使用，用于研究受调控的基因表达和单核苷酸多态。该系统将基于聚合酶链式反应(PCR)的热循环仪与使用激光检测荧光染料的硬件仪器和软件集成在一起。它将为这组研究人员提供一个实时定量核酸序列的系统，在广泛的动态范围内具有准确性和重复性，并具有高通量。通过在单个仪器中结合热循环、荧光检测和软件，该系统能够逐个周期地检测核酸产物的增加。此外，数据输出可以在序列扩增后立即获得，而不需要额外的纯化步骤或凝胶电泳分析。实时定量检测特定的核酸序列是基于在同一探针上加入报告染料和猝灭染料的荧光探针的使用，以及基于Taq DNA聚合酶的5‘核酸酶活性的探针切割。探针裂解将报告染料从猝灭剂中分离出来，从而恢复荧光。该仪器的光谱仪和电荷耦合设备摄像头探测到荧光发射。该系统使用96个样品井平台、一个用于诱导荧光的Ar激光器和一个多路光纤阵列。这些组件包括：(1)内置96孔位置热循环仪的7700序列检测器；(2)Apple Power Macintosh系统，具有16MB RAM、500-Mb硬盘驱动器、120 MHz Power Macintosh系统、1.4-Apple Super Drive软盘驱动器、CD ROM驱动器、15英寸彩色显示器和惠普1600C彩色Deskjet打印机。序列检测软件用于仪器控制、数据收集和自动数据分析。ABI PRISM 7700系统将成为加州大学圣地亚哥分校医学院PHS支持研究的关键核心资源。