INTEGRATED HIGH THROUGHPUT DNA SEQUENCING

集成高通量 DNA 测序

• 批准号: 2674288
• 负责人: STEVAN B JOVANOVICH
• 金额: $ 87.66万
• 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 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2674288
• 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.

用于样品制备和DNA测序的集成系统是 建议。计算机控制的系统将自动执行以下所有步骤 从琼脂平板中挑选重组克隆用于原代生产 序列数据。DNA测序系统将消除所有瓶颈 发生在亚克隆和获得原始序列数据之间， 显著降低了人工和试剂成本，并具有 每台机器每年高达250兆基的原始DNA序列-更多 吞吐量比现有的 技术。DNA测序系统将由四个基本部分组成 硬件组件-克隆选取器、微提取模块(MEM)、 一种处理小容量聚合酶链式反应和循环的装置 测序反应(PCR/CS)和毛细管阵列电泳(CAE) DNA测序仪。DNA测序系统将利用这些能力 既是现有的原型CAE仪器，这是一个高吞吐量 毛细管阵列DNA测序仪和菌落采集器， 功能齐全的高吞吐量设备。此外，系统的主要模块 操作软件已经开发出来了。该项目的目标 1)设计和建造流程型PCR/CS装置；2)设计 并构建了微提取模块，并将其与克隆 Picker，3)评估改进的扩增和测序化学以 简化建议的系统，以及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 协助完成DNA序列采集阶段 人类基因组计划，并使DNA测序应用于 人类遗传病诊断的临床环境和 生物制药的发展。