APPLIED BIOSYSTEMS MODEL 7700 01 SEQUENCE DETECTOR

应用生物系统 7700 01 型序列检测器

• 批准号: 2503055
• 负责人: David Proud
• 金额: $ 9.13万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-05-15 至 1999-05-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2503055
• 关键词: biomedical equipment; biomedical equipment purchase; computer data analysis; computer program /software; polymerase chain reaction

This application requests a Perkin-Elmer ABI Prism 7700-01 Sequence Detector equipped with Primer Express and Sequence Detection software. An additional GeneAmp PCR system 960 and a Macintosh PowerMac 9500 computer with printer are also requested to equip a central facility at the John Hopkins Asthma and Allergy Center to be used by funded investigators from 3 separate divisions that are based at the Center. These investigators will use the Sequence Detector for 1) Real-time, quantitative RT-PCR for a range of cytokines that are involve din inflammatory disease on the airways; 2) to replace radioactive-allele-specific oligonucleotide hybridization for genotype analysis; and 3) to provide a new method for determination of exogenous/endogenous gene copy in transgenic mice. The linear dynamic range of the ABI Prism system covers at least five orders of magnitude and thereby greatly reduces the need for serial dilutions of samples. The system uses an oligonucleotide probe with both a reporter and quencher dye attached, which anneals specifically between forward and reverse primers. The probe is cleaved by the 5' nuclease activity of Taq polymerase to release the reporter dye and generate a sequence-specific signal. Fluorescence intensity correlates directly with a number of amplified chains. Currently, no such system exists anywhere at the John Hopkins Institutions. To maximize efficiency of the proposed facility, all post-acquisition data analysis will be performed using a separate PowerMac 9500 computer system that will also serve as a data storage/server system. The 200-MHz microprocessor will permit rapid data analysis. Finally, an additional GeneAmp 9600 thermal cycle is requested to match the closed-tube assays format and cycle conditions used by the ABI 7700 system. Thus, samples with known sequence variations, such as mutations or polymorphisms currently detected by allele-specific oligonucleotide hybridization, or PCR followed by restriction enzyme digestion will be run on the model 9600 cycler and transferred to the ABI 7700 for probe detection. This will maximize available run-time on the ABI model 7700 for the multiple investigators who require real-time quantitative applications. The availability of this powerful new technology will greatly aid the research of a large group of productive and interactive faculty.

此应用程序请求 Perkin-Elmer ABI Prism 7700-01 序列 检测器配备 Primer Express 和序列检测软件。一个 额外的 GeneAmp PCR 系统 960 和 Macintosh PowerMac 9500 计算机 还要求在约翰配备中央设施 霍普金斯哮喘和过敏中心将由来自以下国家资助的研究人员使用 中心设有 3 个独立的部门。这些调查人员 将使用序列检测器进行 1) 实时定量 RT-PCR 一系列与炎症性疾病有关的细胞因子 航空公司； 2) 取代放射性等位基因特异性寡核苷酸 基因型分析杂交； 3）提供一种新方法 转基因小鼠中外源/内源基因拷贝的测定。 ABI Prism 系统的线性动态范围至少涵盖五个 数量级，从而大大减少了串行的需要 样品的稀释液。该系统使用寡核苷酸探针 附有报告基团和淬灭基团染料，在 正向和反向引物。探针被 5' 核酸酶切割 Taq 聚合酶的活性释放报告染料并产生 序列特异性信号。荧光强度直接与 一些扩增链。目前，任何地方都不存在这样的系统 约翰霍普金斯大学。 为了最大限度地提高拟议设施的效率，所有采集后数据 将使用单独的 PowerMac 9500 计算机系统进行分析 它还将用作数据存储/服务器系统。 200MHz 微处理器将允许快速数据分析。最后，附加一个 GeneAmp 9600 热循环需要匹配闭管检测 ABI 7700 系统使用的格式和循环条件。因此，样品 具有已知的序列变异，例如突变或多态性 目前通过等位基因特异性寡核苷酸杂交检测到，或 PCR 和限制酶消化将在 9600 型上运行 循环仪并转移至 ABI 7700 进行探针检测。这将 最大化 ABI 型号 7700 上的多个可用运行时间 需要实时定量应用的研究人员。 这项强大的新技术的出现将极大地帮助 一大群富有成效和互动的教师的研究。