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.

提出了一种加速毛细管阵列DNA测序系统的开发和实现的方案。 该系统将包括与分离基质、试剂和客户支持结构偶联的384毛细管仪器，以在小于或等于2小时内实现大于1200个碱基的一致DNA测序读数。 该系统将有助于将现有的最先进的长读序测序技术整合到生产设施中。 该系统将以比现有毛细管系统快约10倍的吞吐量产生数据，并将试剂使用量减少约20倍。DNA测序仪将包括一个4-PMT共聚焦扫描仪设计与4个96毛细管阵列恒温在固态加热元件，操作高达85摄氏度。分离矩阵将基于东北大学开发的高分子量和低分子量线性聚丙烯酰胺的混合物，该混合物已被证明可提供1200个碱基的高质量序列数据。 样品制备将在384毛细管试剂卡中进行，该试剂卡以500 nl规模制备样品。 这些组件利用了东北大学、分子动力学和阿默舍姆法玛西亚的现有技术，但协同工作将使系统的开发具有显著增强的性能。该项目的目标是：1）设计和组装仪器，2）制造优化的试剂，3）改进现有的软件，4）提供样品制备设备和方案，5）提供现场支持。这一新系统的加速开发将大大促进人类基因组计划的DNA序列获取阶段和其他序列密集型工作。