INTEGRATED DNA SEQUENCING SYSTEM

集成 DNA 测序系统

• 批准号: 6155142
• 负责人: STEVAN B JOVANOVICH
• 金额: $ 53.59万
• 依托单位: MOLECULAR DYNAMICS, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-06-20 至 2002-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6155142
• 关键词: biomedical equipment development; biotechnology; computer assisted sequence analysis; high throughput technology; microarray technology; nanotechnology; nucleic acid sequence

DESCRIPTION: (Applicant's Abstract) The Human Genome Project is rapidly developing a draft consensus sequence of the Human Genome. The immense value of this information has triggered the scale up of production rates and the incorporation of high-throughput capillary-array electrophoresis instruments. Further improvements are needed to decrease sample costs, increase automation, and improve throughput rates. This project will create an automated, integrated DNA analysis system that prepares nanoliter (nL)-scale sequencing reactions and analyzes the reactions in an ultrahigh-throughput, microchip-based DNA sequencer. The integrated system will consume about one-hundredth of the reagents used today and have a ten-fold higher throughput rate. The integrated system will consist of two main modules, a sample preparation module and an ultrahigh-throughput DNA analysis module. The sample preparation module will use arrays of 384 capillaries in \cassettes to move liquids and perform biochemical reactions in 125 nL volumes. This module will automate shotgun sequencing beginning with lysed clones to produce fluorescently labeled cycle sequencing reactions for capillary or microchip analysis. The microchip-based DNA analysis module will load, run, and detect DNA samples in microchannels for sequencing and genotyping. This module will use robotics to transfer samples from microtiter plates to sample loading ports on a microchip, dock the microchip with electrodes under a detector, inject samples and separate them. The two modules will be developed simultaneously and then integrated into a complete automated sequencing system. The project will develop a series of prototypes that will be alpha-tested in the Molecular Dynamics Production Sequencing Group and then beta-tested at our collaborators at the Joint Genome Institute, the Washington University Genome Sequencing Center, and the Macromolecular Analysis Facility at the University of Wisconsin-Madison. The results of the external testing will be used to verify the system performance, refine the prototypes, and streamline the integration of this new technology into genome centers and core facilities.

描述：(申请者摘要)人类基因组计划进展迅速 开发人类基因组的共识序列草案。它的巨大价值 这一信息引发了生产率的扩大和 采用高通量毛细管阵列电泳仪。 需要进一步改进，以降低样品成本，提高自动化程度， 并提高吞吐量。 该项目将创建一个自动化的、集成的DNA分析系统 准备纳升(NL)规模的排序反应并分析反应 在超高通量、基于微芯片的DNA测序仪中。集成的 系统将消耗目前使用的试剂的百分之一，并具有 吞吐速度提高十倍。综合系统将由两个主要部分组成 模块、样品制备模块和超高通量DNA分析 模块。样品制备模块将使用384个毛细管阵列 用于移动液体并在125毫升体积内进行生化反应的盒。 本模块将从裂解克隆开始自动进行鸟枪测序，以 产生荧光标记的循环测序反应用于毛细管或 微芯片分析。基于微芯片的DNA分析模块将加载、运行和 检测微通道中的DNA样本以进行测序和基因分型。本模块 将使用机器人将样本从微量滴定板转移到样本装载 微芯片上的端口，将微芯片与检测器下的电极对接， 注射样本并将其分离。将开发这两个模块 同时，然后集成到一个完整的自动测序系统中。 该项目将开发一系列原型，这些原型将在 分子动力学产品测序小组，然后在我们的 华盛顿大学联合基因组研究所的合作者 测序中心和大学的大分子分析设施 威斯康星州麦迪逊分校。外部测试的结果将用于 验证系统性能，改进原型，并简化 将这项新技术整合到基因组中心和核心设施中。