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INTEGRATED HIGH THROUGHPUT DNA SEQUENCING

集成高通量 DNA 测序

基本信息

批准号：
6136596
负责人：
STEVAN B JOVANOVICH
金额：
$ 14.61万
依托单位：
MOLECULAR DYNAMICS, INC.
依托单位国家：
美国
项目类别：
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

项目摘要

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.

一种用于样品制备和DNA测序的集成系统，提出了计算机控制系统将自动化所有步骤，从琼脂平板上挑取重组克隆，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，显著降低劳动力和试剂成本，并具有生产能力每台机器每年可处理高达250兆字节的原始DNA序列-更多吞吐量的数量级增长，技术. DNA测序系统将包括四个基本的硬件组件-克隆拾取器、微萃取模块（MEM）、用于进行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测序系统将利用现有的原型CAE仪器，这是一个高通量毛细管阵列DNA测序仪和菌落挑选器，功能性高通量装置。此外，操作软件已经开发出来了。该项目的目标设计并构建了一套流式PCR/CS装置，构建一个微萃取模块，挑选器，3）评估改进的扩增和测序化学，简化所提出的系统，以及4）整合化学和硬件并在大规模测序中测试DNA测序系统项目集成DNA测序系统的开发将促进完成的DNA序列采集阶段的人类基因组计划，并使DNA测序的应用，诊断人类遗传性疾病的临床环境，生物制药的发展。