AUTOMATED DNA SEQUENCING INSTRUMENTATION

我们请求资助用于自动化DNA的仪器 测序 我们已经确定一个集中的测序设施 将为19个主要用户节省大量时间和消耗品成本 以及加州理工学院化学和生物系的其他人，而在 同时为项目的成功提供关键数据。 在日常项目中进行了大量的排序， 这一切都必须通过人工方法来完成， 个别学生，博士后研究员和技术人员，其中许多人 没有DNA测序的经验。 这导致缺乏 效率和竞争力的丧失。 我们要三样东西 相关设备：自动DNA测序仪， 电泳凝胶，一个机器人站进行聚合 DNA测序技术所基于的反应，以及计算机 更新现有测序分析设备的硬件和软件 以使其能够科普将增加的负荷， 自动测序设备的出现 自动化测序 基于荧光标记的方法至少有四个优于人工的优点 基于放射性的方法：节省消耗品成本，节省劳动力， 安全，最重要的是，结果的质量。 也许最有用的 基于自动化方法的中心测序设施的方面是 自动化序列可以与计算机集成的程度 系统. 因此，该设施将提高生产力和质量 主要用户的研究。 这里记录了19个主要用户， 人所提及 实验室容纳了300多名研究人员，学生， 博士后研究员和技术人员。 的主要应用 仪器将是涉及广泛克隆的项目， 新基因的测序和/或基因工程的构建 需要序列分析的构建体。 这些项目包括， 其他的，RNA剪接相关基因的测序，分子生物学 控制神经嵴发育，在存活和分化 果蝇神经元的功能，在膜兴奋性， 在果蝇的细胞周期中控制体节的发育 调控，酵母中的DNA复制，细胞命运和形态发生 在几个有机体中。 其中一个项目涉及对大量的 随机选择的cDNA克隆参与花发育， 阿拉伯茶

We request funding for instrumentation to be used in automated DNA sequencing. We have determined that a centralized sequencing facility will save considerable time and consumable costs for the 19 major users and others in the chemistry and biology divisions at Caltech, while at the same time contributing data crucial to the success of the projects. Large amounts of sequencing are carried out in the day to day projects, and it all must be done by manual methods based on radioactivity by individual students, post-doctoral fellows and technicians, many of whom have no previous experience in DNA sequencing. This leads to lack of efficiency and loss of competitiveness. We are requesting three items of related equipment: an automated DNA sequencer that runs and reads electrophoresis gels, a robotic station to carry out the polymerization reactions on which DNA sequencing technology is based, and computer hardware and software to update an existing Sequencing Analysis Facility to make it able to cope with the increased load that will arise with the advent of the automated sequencing facility here. Automated sequencing based on fluorescent labels has at least four advantages over manual methods based on radioactivity: saving in consumable costs, labor saving, safety, and, most important, quality of results. Perhaps the most useful aspect of a central sequencing facility based on automated methods is the extent to which the automated sequence can be integrated with computer systems. The facility will, thus, enhance the productivity and quality of the studies of the major users. There are 19 major users documented here and additional users are mentioned. The laboratories houses over 300 investigators, students, post-doctoral fellows and technicians. The major applications of the instruments will be projects that involve extensive cloning and sequencing of novel genes and/or construction of genetically engineered constructs that require sequence analysis. The projects cover, among others, sequencing of genes involved in RNA splicing, in molecular control of neural crest development, in the survival and differentiated function of neurons in Drosophila, in membrane excitability, in controlling body segment development in Drosophila, in cell cycle regulation, in DNA replication in yeast, in cell fate and morphogenesis in several organisms. One project involves sequencing large numbers of randomly-chosen cDNA clones involved in floral development in Arabidipsis.