LOW VISCOSITY SEPARATION MEDIA FOR DNA SEQUENCING

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.