AUTOMATED SEQUENCING SYSTEM FOR THE HUMAN GENOME PROJECT

人类基因组计划自动测序系统

• 批准号: 2415452
• 负责人: Gregory T. Went
• 金额: $ 161.23万
• 依托单位: CURAGEN CORPORATION
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-05-01 至 1999-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2415452
• 关键词: DNA; charge coupled device camera; computer assisted sequence analysis; computer program /software; electrophoresis; fluorescent dye /probe; gel; gel electrophoresis; genetic mapping; genome; human genetic material tag; nucleic acid sequence; nucleobase; optics; robotics

The genetic information encoded in the human genome specifies all life processes from conception to death, and contains within it the origin of over 3500 known genetic diseases. It is necessary to substantially reduce the cost and dramatically raise the speed of producing accurate DNA sequences in order to determine the sequence of the 3 billion basepairs of DNA that compose the human genome. This proposal describes a three-year project to develop, integrate and distribute into parallel sequencing projects a fluorescent sequencing instrument and software system capable of meeting these logistical and cost demands. This approach allows for the identification and removal of the major bottlenecks encountered in producing a fully automated system, without duplication of mapping, sample processing or informatics efforts ongoing at other HGP facilities. Current bottlenecks arise primarily from inefficiencies in workflow and data analysis. Specifically, we will develop and integrate into existing centers the following: 8-dye bi-directional sequencing methods; solid phase loading of an advanced electrophoresis instrument; high-throughput and low cost ready-to-run disposable microreplicated separation sub- systems; sensitive detection optics; and advanced base-calling software. These advances will enable the completion of fully automated systems including all the steps necessary to proceed from a physical map to a finished sequence with a quantifiable measure of confidence. Full process automation coupled with automated data analysis and reduced materials cost will enable a 60-fold increase in throughput with the required 10-fold decrease in cost relative to systems based on currently available technology. This project is coordinated by CuraGen Corporation and builds on the automation, data analysis, sequencing instrumentation, and fabrication experience of collaborators at Soane BioSciences, MIT. The final product of our efforts will be tested and ready for wide-spread distribution for use in sequencing the human genome.

人类基因组中编码的遗传信息指定了所有生命 从构思到死亡的过程，其中包含 超过3500种已知遗传疾病。 有必要大大减少 成本并大大提高了产生准确DNA的速度 序列以确定30亿底座的序列 组成人基因组的DNA。 该提案描述了三年 项目开发，集成和分布到并行测序 项目荧光测序仪器和软件系统 满足这些后勤和成本要求。 这种方法允许 在 生产一个全自动系统，而无需重复映射，样品 其他HGP设施正在进行的处理或信息学工作。 当前的瓶颈主要来自工作流程中的低效率和 数据分析。 具体而言，我们将开发并集成到现有 中心以下：8-DYE双向测序方法；坚硬的 高级电泳仪器的相位载荷；高通量 和低成本即时运行的一次性微积分离子 - 系统；敏感检测光学元件；和先进的基础呼叫软件。 这些进步将使完全自动化系统的完成 包括从物理地图到一个的所有必要步骤 完成的序列具有可量化的置信度。 完整的过程 自动化以及自动数据分析和降低的材料成本 所需的10倍将使吞吐量增加60倍 基于当前可用的系统相对于系统的成本降低 技术。 该项目由Curagen Corporation协调​​，并建立 关于自动化，数据分析，测序仪器和 MIT Soane Biosciences的合作者的制造经验。 这 我们的努力的最终产品将经过测试，并准备好进行广泛 用于测序人基因组的分布。