INTEGRATED HIGH THROUGHPUT DNA SEQUENCING

用于样品制备和 DNA 测序的集成系统 提议。 计算机控制系统将自动执行所有步骤 从琼脂平板中挑选重组克隆以生产初级产品 序列数据。 DNA测序系统将消除所有瓶颈 发生在亚克隆和获取原始序列数据之间， 显着降低劳动力和试剂成本，并具有吞吐量 每台机器每年最多可处理 250 兆碱基的原始 DNA 序列 - 更多 与现有的吞吐量相比，吞吐量增加了一个数量级 技术。 DNA测序系统将由四个基本部分组成 硬件组件 - 克隆选择器、微提取模块 (MEM)、 用于处理小体积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 测序系统将利用以下功能 现有原型 CAE 仪器，这是一种高通量 毛细管阵列DNA测序仪和菌落选择器， 功能性高通量设备。此外，主要模块 操作软件已经开发出来。 项目的目标 目的是：1) 设计和构建流程 PCR/CS 装置，2) 设计 构建微提取模块并与克隆集成 选择器，3) 评估改进的扩增和测序化学 简化所提出的系统，并且 4) 集成化学和硬件 并在大规模测序中测试DNA测序系统 项目。 集成DNA测序系统的开发将 促进DNA序列采集阶段的完成 人类基因组计划并使 DNA 测序的应用成为可能 人类遗传疾病诊断的临床环境和 生物制药的发展。

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.