INTEGRATED DNA SEQUENCING SYSTEM

集成 DNA 测序系统

• 批准号: 6388348
• 负责人: STEVAN B JOVANOVICH
• 金额: $ 51.34万
• 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 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6388348
• 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：（申请人摘要）人类基因组计划正在迅速推进 制定人类基因组共识序列草案。的巨大价值 这一信息引发了生产率的提高和 结合高通量毛细管阵列电泳仪器。 需要进一步改进以降低样品成本、提高自动化程度、 并提高吞吐率。 该项目将创建一个自动化、集成的 DNA 分析系统 准备纳升 (nL) 规模的测序反应并分析反应 在超高通量、基于微芯片的 DNA 测序仪中。集成的 系统将消耗当今使用的试剂的大约百分之一，并且具有 吞吐率提高十倍。该集成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 \盒用于移动液体并在 125 nL 体积中进行生化反应。 该模块将从裂解的克隆开始自动进行鸟枪测序 产生毛细管荧光标记的循环测序反应或 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。该模块 将使用机器人技术将样品从微量滴定板转移到样品装载 微芯片上的端口，将微芯片与探测器下方的电极对接， 注入样品并分离它们。将开发两个模块 同时进行，然后集成到完整的自动化测序系统中。 该项目将开发一系列原型，并将在 2019 年进行 alpha 测试。 分子动力学生产测序小组，然后在我们的进行 beta 测试 华盛顿大学基因组联合基因组研究所的合作者 大学测序中心和高分子分析设施 威斯康星-麦迪逊。外部测试的结果将用于 验证系统性能，完善原型并简化 将这项新技术整合到基因组中心和核心设施中。