ABI PRISM 7700 SEQUENCE DETECTION SYSTEM

传统的基因剂量和表达分析策略，如半定量的Southern和northern blot分析，已经被更高效的基于聚合酶链反应（PCR）的序列多态性检测所取代。使用标准PCR协议，加上逆转录酶进行RNA分析，提高了速度，吞吐量和易用性，可以实现半定量终点数据分析。然而，PCR产物的终点分析用于定量测定是有限的，因为它不代表过程的线性阶段。终点PCR分析的有用性因引入PCR后变异而进一步复杂化，包括样品处理和视觉数据收集错误。所有这些考虑使得从终点PCR分析中获得准确的定量数据在技术上变得困难。应用生物系统公司（ABI） PRISM 7700序列检测系统等仪器提高了基因和基因转录物定量序列检测的效率、速度和范围。由于PRISM 7700可重复的实时荧光数据收集，可以进行比较和直接定量分析。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，可以更快速和更经济有效地进行。杰克逊实验室（TJL）因其在哺乳动物遗传学方面的科学贡献而得到公认。TJL大约40%的研究项目使用半定量DNA和RNA分析技术。仅从节省的成本来看，该系统的可用性将使这些当前项目的吞吐量增加34%。我们三分之一（11）的工作人员的项目，由16个NIH拨款资助，将通过使用该仪器大大提高。未来项目的范围也将扩大，从而大大提高该机构科学工作人员的整体生产力。使TJL工作人员能够更迅速地筛选、鉴定和提供新的人类疾病模型，将有利于科学界。申请资金用于购买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。

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.