IMPLEMENTATION OF A 384 CAPILLARY SEQUENCING SYSTEM

384 毛细管测序系统的实施

• 批准号: 6181851
• 负责人: STEVAN B JOVANOVICH
• 金额: $ 116.36万
• 依托单位: MOLECULAR DYNAMICS, INC.
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6181851
• 关键词: biomedical resource; capillary electrophoresis; chemical synthesis; computer program /software; genetic techniques; genome; nucleic acid sequence; reagent /indicator; technology /technique development

An accelerated program for the development and implementation of a capillary-array DNA sequencing system is proposed. The system will comprise a 384-capillary instrument coupled with the separation matrix, reagents, and customer support structure necessary to achieve consistent DNA sequencing reads greater than 1200 bases in less than or equal to 2 hours. The system will facilitate integration of extant state-of-the-art technologies for long-read sequencing into production facilities. The system will produce data with a throughput approximately 10x faster than existing capillary-based systems, and reduce reagent usage by approximately 20x. The DNA sequencing instrument will consist of a 4-PMT confocal scanner design with 4 96-capillary arrays thermostated in a solid-state heating element, operating as high as 85 degree C. The separation matrix will be based on mixtures of high- and low-molecular weight linear polyacrylamide developed at Northeastern University that have already been shown to give high-quality sequence data to 1200 bases. Sample preparation will be done in a 384-capillary cassette that prepares samples on a 500nl scale. These components leverage existing technologies at Northeastern University, Molecular Dynamics, and Amersham Pharmacia, but in concert will enable development of a system with dramatically enhanced performance. The goals of the project are to: 1) Design and assemble the instrumentation, 2) manufacture optimized reagents, 3) improve on available software, 4) deliver sample preparation devices and protocols, 5) deliver field support. Accelerated development of this new system will greatly facilitate the DNA sequence acquisition phases of the Human Genome Project and other sequence-intensive efforts.

提出了用于开发和实施毛细管阵列DNA测序系统的加速程序。 该系统将组成384个毛细管仪器，并结合分离矩阵，试剂和客户支持结构，以实现一致的DNA测序读数读数在小于或等于2小时内大于1200个基础。 该系统将促进将现有的最新技术集成，以进行长阅读测序与生产设施。 该系统将比现有的基于毛细管的系统快10倍生产数据，并将试剂使用率减少约20倍。 DNA测序仪器将由4-PMT共聚焦扫描仪设计组成，其在固态加热元件中恒温的4 96个毛细管阵列，运行高达85度C。分离矩阵将基于高和低的混合物。 - 在东北大学开发的分子量线性聚丙烯酰胺，这些聚丙烯酰胺已被证明可为1200个基碱基提供高质量的序列数据。 样品制备将以384个毛细血管盒进行500NL尺度制备样品。 这些组件利用了东北大学，分子动力学和Amersham Pharmacia的现有技术，但共同开发具有大幅增强性能的系统。该项目的目标是：1）设计和组装仪器，2）制造优化的试剂，3）改进可用软件，4）提供样品准备设备和协议，5）提供现场支持。该新系统的加速发展将极大地促进人类基因组项目和其他序列密集型努力的DNA序列采集阶段。