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.
