A MULTI-USER GENE ARRAYER CORE FACILITY

在生物学中使用DNA阵列将对我们对形状和形式、癌症生物发生、基因回路、药物开发和进化等基本问题的理解产生深远的影响。这些阵列的一个主要用途是分析信使RNA水平的时空变化，其中可以同时观察对照和实验样品。最近对各种环境条件下酵母生命周期的全基因组分析，对大肠杆菌和Caulobacter的类似研究，最近对不同b细胞淋巴瘤的分析，以及恶性疟原虫的表达谱都是令人惊叹的例子，表明微阵列分析将使我们能够一次研究的不仅仅是几个基因，而是整个生物体的遗传补充。这一信息具有不可估量的价值，特别是当它与其他遗传方法相结合时。它将使我们不需要对生命周期中使用的东西进行分类，但它也将使我们能够在全基因组的基础上提出问题，了解密切相关物种之间的差异，发育期间和病毒感染后的基因调控，以及肿瘤发生过程中的基因调控。为了最佳地使用基因阵列技术，特别是涉及几百个基因的阵列，需要三个集成工作站，阵列本身，解读阵列上信号的阅读器，以及在所需的大规模上准备和分析DNA模板的机器人系统。本建议为其中第三个项目寻求资源。提出了一种能够重新格式化，复制，纯化和分析超过20,000个不同DNA样本的机器人工作站。它将并入一个核心多用户阵列设施，由训练有素的技术人员监督，他们的职责将是管理设备和培训最终用户。微阵列和读取器已经在这个设施中建造，现在正在使用中。所提出的机器人将与阵列和读取器集成。

The use of DNA arrays in biology will have a profound influence on our understanding of such basic questions as shape and form, cancer biogenesis, gene circuits, drug development, and evolution. One major use of these arrays is to analyze temporal and spatial variation in messenger RNA levels where control and experimental samples can be observed simultaneously. The recent genome-wide analysis of the yeast life cycle under a variety of environmental conditions, similar studies in E. coli and Caulobacter, the recent analysis of different B-cell lymphomas, and the expression profiling of Falciparum are all stunning examples of how micro array analysis will allow us to study not just a few genes at a time, but the entire genetic complement of an organism. This information is of inestimable value, especially when it is combined with other genetical approaches. It will allow us not to catalogue what is used during a life cycle, but it will also low us to ask questions on a genome-wide basis about differences between closely related species, gene regulation during development and after viral infection, and during the progress of tumorigenesis. To use gene array techniques optimally, especially for arrays involving more than a few hundred genes, three integrated work-stations are required, the arrayed itself, a reader to interpret signals on the array, and a robotic system to prepare and analyze DNA templates on the required large scale. This proposal seeks resources for the third of these items. A robotic work-station capable of reformatting, replicating, purifying, and analyzing upwards of 20,000 different DNA samples is proposed. It will be integrated into a Core Multi-user Array Facility supervised by trained technical staff whose responsibilities will be to manage the equipment and train the end-user. The micro arrayed and reader have been constructed in this facility and are now in use. The proposed robotics will be integrated with the arrayed and reader.