Automated high-throughput analysis system

描述(由申请人提供)：该提案是为索尔克研究所仪器核心的自动化高通量分析系统获得资金。该仪器系统将包括一个Eppendorf epMotion 5075 VAC-TMX高通量DNA制备工作站，一个用于液体分配和分析的Eppendorf epMotion 5075 LH液体处理站，以及一个用于各种基于细胞和体外分析的分子设备FlexStation 3多模式检测器。所需的高通量分析系统是对索尔克研究所强大的化学库筛选核心的补充。我们将利用这个系统来建立和使用基因组学筛选的cDNA和shRNA试剂文库。高通量的基因组DNA纯化系统将允许为基于细胞的分析准备cDNA库或shDNA构建库。液体处理站将把cDNA、shDNA和慢病毒颗粒分配到哺乳动物细胞中。多模探测器平板读取器将测量吸光度、荧光强度、发光、荧光偏振和时间分辨荧光。Flexstation 3平板阅读器将使索尔克研究所的研究人员能够使用基于荧光的染料以高通量方式测量细胞内钙水平或膜电位的快速变化。这些仪器的组合将为使用基因组学方法提供一个强大而灵活的平台，以加快对多个不同实验室的生物过程的理解。我们已经确定了10个用户实验室，用于立即使用该系统进行各种分析开发、筛查和筛查后跟进。拟议的仪器系统将与索尔克研究所现有的仪器核心集成在一起。一个核心仪器使用委员会将确保仪器系统由训练有素的人员操作和维护。 与公共健康相关：该提案旨在为索尔克研究所的机器人系统获得资金。这一仪器系统将使研究人员能够以高度的准确性、重复性和速度对数千个生物样本进行筛选，以了解基因和蛋白质在健康和患病细胞中的功能。

DESCRIPTION (provided by applicant): The proposal is to obtain funds for an automated high throughput analysis system for the instruments core at the Salk Institute. The instrument system will comprise of an Eppendorf epMotion 5075 VAC-TMX high throughput DNA preparation workstation, an Eppendorf epMotion 5075 LH liquid handling station for liquid dispensing and assay set up, and a Molecular Devices FlexStation 3 multi- mode detector for various cell based and in vitro assays. The requested high throughput analyses system is needed to complement the strong chemical library screening core at the Salk Institute. We will use this system to generate and use libraries of cDNA and shRNA reagents for genomics screening. The high throughput genomic DNA purification system will allow preparation of libraries of cDNA or shDNA constructs for cell based assays. The liquid handling station will dispense the cDNA, shDNA and lentiviral particles into mammalian cells. The multimode detector plate reader will measure absorbance, fluorescence intensity, luminescence, fluorescence polarization and time-resolved fluorescence. The Flexstation 3 plate reader will enable researchers at the Salk Institute to use fluorescence based dyes to measure rapid changes in intracellular calcium level or of membrane potential in a high throughput manner. The combination of these instruments will offer a powerful yet flexible platform for use of genomics approaches to accelerate understanding of biological processes in multiple diverse laboratories. We have identified 10 user laboratories for immediate use of the system for various assay development, screening and post-screening follow-up. The proposed instrument system will be integrated to the existing Salk Institute instrument cores. A core instrument use committee will ensure that the instrument system is operated and maintained by trained personnel. PUBLIC HEALTH RELEVANCE: The proposal is to obtain funds for a robotic system at the Salk Institute. This instrument system will enable researchers to screen through thousands of biological samples with high degree of accuracy, reproducibility and speed to understand how genes and proteins function in healthy and diseased cells.