Current high-throughput DNA sequencing systems can produce sequence data at a theoretical rate of approximately 29 megabases per year. Capillary Array Electrophoresis (CAB) systems can increase the DNA sequencing throughput by an order-of-magnitude over currently available systems. A flowable separation medium is one of the key components of the CAB system, and improvements in separation media significantly lower the cost of a CAB system. The moderate-to-high viscosities of the separation media used by previous researchers makes it a formidable task to design a pressurized capillary refilling station for a commercial CAB system. The goal of Phase I of this proposal is to develop low-viscosity media which will enable single base resolution out to 1000 bases. Using low-viscosity separation media will simplify the pressure-vessel design issues of the CAB system, resulting in a substantial reduction in cost. Use of such high-throughput CAB systems will revolutionize large-scale sequencing projects, and substantially reduce the cost per base. Phase II studies will extend the utility of low-viscosity and high-resolution media with less expensive dynamically-coated capillaries by eliminating the need for high-cost covalently-coated capillaries. This will further reduce the cost per system and per base sequences.
A CAE system has significant potential to improve the U.S. economic growth and productivity by accelerating the rate that new genetic information is generated. Reliable, cost-efficient sequencing and genetic typing will have significant impact on Human Genome Project, healthcare, agribusiness, forensic analysis, and pharmaceutical industry.
| Dolnik, V; Gurske, W A (1999) Capillary electrophoresis in sieving matrices: selectivity per base, mobility slope, and inflection slope. Electrophoresis 20:3373-80 |
