INTEGRATED HIGH THROUGHPUT DNA SEQUENCING

集成高通量 DNA 测序

• 批准号: 2562669
• 负责人: STEVAN B JOVANOVICH
• 金额: $ 116.48万
• 依托单位: MOLECULAR DYNAMICS, INC.
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-07-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2562669
• 关键词: biomedical automation; capillary electrophoresis; method development; molecular cloning; nucleic acid sequence; polymerase chain reaction

An integrated system for sample preparation and DNA sequencing is proposed. The computer controlled system will automate all steps from picking recombinant clones from agar plates to production of primary sequence data. The DNA sequencing system will eliminate all bottle necks that occur between subcloning and obtaining raw sequence data, significantly reduce both labor and reagent costs, and have a throughput of up to 250 megabases of raw DNA sequence per machine per year - more than an order-of-magnitude increase in throughput over existing technologies. The DNA sequencing system will consist of four basic hardware components - a clone picker, a microextraction module (MEM), a device for processing small volume polymerase chain reaction and cycle sequencing reactions (PCR/CS), and a capillary array electrophoresis (CAE) DNA sequencer. The DNA sequencing system will leverage the capabilities of both an existing prototype CAE instrument, which is a high throughput capillary array DNA sequencing instrument, and a colony picker, which is a functional high throughput device. In addition, major modules of the operating software have already been developed. The goals of the project are to: 1) design and construct a flow process PCR/CS device, 2) design and construct a micro-extraction module and integrate it with the clone picker, 3) evaluate improved amplification and sequencing chemistry to simplify the proposed system, and 4) integrate the chemistry and hardware and test the DNA sequencing system within a large-scale sequencing project. Development of the integrated DNA sequencing system will facilitate the completion of the DNA sequence acquisition phase of the Human Genome Project and enable the application of DNA sequencing to clinical settings for the diagnosis of human genetic diseases and the development of biopharmaceuticals.

样品制备和DNA测序的集成系统是 建议的。 计算机控制的系统将自动化所有步骤 从琼脂板上挑选重组克隆到主要的生产 序列数据。 DNA测序系统将消除所有瓶颈 在亚克隆和获得原始序列数据之间发生这种情况， 显着降低劳动力和试剂成本，并具有吞吐量 每年最多每台机器的原始DNA序列250兆瓦 - 更多 比现有的吞吐量增加了 技术。 DNA测序系统将由四个基本组成 硬件组件 - 克隆拾取器，微萃取模块（mem），一个 处理小体积聚合酶链反应和循环的设备 测序反应（PCR/CS）和毛细管阵列电泳（CAE） DNA测序仪。 DNA测序系统将利用功能 两种现有的原型CAE仪器，这是高通量的 毛细管阵列DNA测序仪器和一个菌落选择器， 功能性高通量设备。此外， 操作软件已经开发。 项目的目标 为：1）设计和构建流程PCR/CS设备，2）设计 并构建一个微萃取模块并将其与克隆集成 选择器，3）评估改进的扩增和测序化学 简化提出的系统，4）整合化学和硬件 并在大规模测序中测试DNA测序系统 项目。 综合DNA测序系统的开发将 促进完成DNA序列采集阶段的完成 人类基因组项目，并使DNA测序应用于 诊断人类遗传疾病的临床环境和 生物制药的发展。