ABI PRISM 7700 SEQUENCE DETECTION SYSTEM

ABI PRISM 7700 序列检测系统

• 批准号: 2803469
• 负责人: Derry Charles Roopenian
• 金额: $ 10.2万
• 依托单位: JACKSON LABORATORY
• 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/2803469
• 关键词: biomedical equipment; biomedical equipment purchase; nucleic acid quantitation /detection; nucleic acid sequence; polymerase chain reaction

Conventional gene dosage and expression analysis strategies such as semi-quantitative Southern and northern blot analysis have been supplanted by more efficient polymerase chain reaction (PCR) based sequence polymorphism detection. Use of standard PCR protocols, coupled with reverse transcriptase for RNA analysis, has advanced the speed, throughput, and ease with which semi-quantitative endpoint data analysis can be achieved. However, endpoint analysis of PCR products is of limited use for quantitative determinations because it does not represent the linear phase of the process. The usefulness of endpoint PCR analysis is further complicated by the introduction of post PCR variations, including sample handling and visual data collection errors. All of these considerations make it technically difficult to derive accurate quantitative data from endpoint PCR analysis. Instruments such as the Applied Biosystems (ABI) PRISM 7700 Sequence Detection System have increased the efficiency, speed and scope of sequence detection for quantitation of genes and gene transcripts. Comparative and direct quantitative analyses are possible due to the PRISM 7700's reproducible real time fluorescence data collection. This instrument would facilitate common applications including genotyping and transgene copy number/gene dosage detection. More difficult procedures, including pathogen load determination and quantitative gene expression analysis, could be carried out more rapidly and cost effectively. The Jackson Laboratory (TJL) is well recognized for its scientific contributions in mammalian genetics. Approximately 40% of research projects at TJL utilize semi-quantitative DNA and RNA analysis techniques. Based on the cost savings alone, throughput of these current projects would be increased 34% with the availability of this system. Projects of one third (11) of our staff members, funded by 16 NIH grants would be greatly enhanced by access to this instrumentation. The scope of future projects would also be broadened, significantly improving the overall productivity of this institution's scientific staff. The scientific community would be served by enabling TJL staff to screen, characterize and make available new models for human diseases available more rapidly. Funds are requested to purchase the ABI PRISM 7700 Sequence Detection System, necessary computer hardware, operating/data analysis software and uninterruptible power supply.

传统的基因剂量和表达分析策略，如半定量Southern和Northern印迹分析，已经被更有效的基于聚合酶链式反应(PCR)的序列多态检测所取代。使用标准的聚合酶链式反应和逆转录酶进行RNA分析，提高了半定量终点数据分析的速度、吞吐量和简便性。然而，聚合酶链式反应产物的终点分析对定量测定的使用有限，因为它不代表过程的线性阶段。由于引入了聚合酶链式反应后变异，包括样品处理和可视化数据收集错误，终点聚合酶链式反应分析的有用性进一步复杂化。所有这些考虑使得从终点聚合酶链式反应分析中获得准确的定量数据在技术上是困难的。应用生物系统(ABI)PRISM 7700序列检测系统等仪器提高了对基因和基因转录本进行定量的序列检测的效率、速度和范围。由于PRISM7700‘S可重复实时荧光数据采集，可以进行比较和直接的定量分析。该仪器将促进包括基因分型和转基因拷贝数/基因剂量检测在内的常见应用。更困难的程序，包括病原体负荷测定和定量基因表达分析，可以更快、更具成本效益地进行。杰克逊实验室(TJL)在哺乳动物遗传学方面的科学贡献是公认的。TJL大约40%的研究项目使用了半定量DNA和RNA分析技术。单从节省的成本来看，如果这个系统投入使用，目前这些项目的吞吐量将增加34%。我们三分之一(11)的工作人员的项目，由16个NIH赠款资助，通过使用这种仪器将大大增强。未来项目的范围也将扩大，显著提高该机构科学工作人员的整体生产率。科学界将得到帮助，使TJL工作人员能够更快地筛选、确定人类疾病的新模型并提供可用的新模型。需要资金购买ABI PRISM 7700序列检测系统、必要的计算机硬件、操作/数据分析软件和不间断电源。