A MULTI-USER GENE ARRAYER CORE FACILITY

DNA阵列在生物学中的应用将深刻影响我们对形状和形式、癌症生物发生、基因电路、药物开发和进化等基本问题的理解。这些阵列的一个主要用途是分析信使RNA水平的时间和空间变化，其中可以同时观察对照和实验样本。最近对各种环境条件下的酵母生命周期进行的全基因组分析，在大肠杆菌和铜绿假单胞菌中的类似研究，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，都是令人震惊的例子，说明微阵列分析如何使我们能够一次不仅研究几个基因，而且研究一个有机体的整个遗传组成。这些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。它将允许我们不对生命周期中使用的东西进行分类，但它也将降低我们在全基因组基础上询问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形成过程中的基因调控的可能性。为了以最佳方式使用基因阵列技术，特别是对于涉及数百多个基因的阵列，需要三个集成的工作站，阵列本身、用于解释阵列上的信号的阅读器，以及用于大规模准备和分析DNA模板的机器人系统。该提案为其中第三个项目寻求资源。提出了一个能够对20,000多个不同的DNA样本进行重新格式化、复制、纯化和分析的机器人工作站。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，他们的职责是管理设备和培训最终用户。微型阵列和阅读器已在该设施中建造并投入使用。拟议的机器人将与阵列和阅读器集成在一起。

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.