High-field DNA electrophoresis in multiplexed capillaries and microfabricated channels has the potential to provide the order-of- magnitude increases in speed and throughput which are required to sequence the human genome within prescribed time and funding limits. However, advances in separation media are critically needed to enable the practical implementation of these 'microchannel' systems. In Phase I, we have demonstrated the viability of novel, 'thermally-switchable' media which provide the necessary combination of low-viscosity loading and high-resolution separations. In Phase II, we will optimize these matrices to obtain the needed 500+ base read lengths. We will also develop a temperature-ramping method for further increasing read length, and establish protocols for reliable matrix replacement in high-density capillary arrays. Standard procedures will be developed for the polymerization, purification, and processing of our separation media to prepare for the commercialization of this separation media.
The replacement gels and capillaries developed in this research will have immediate commercial utility in capillary electrophoresis instruments. This project will contribute substantially to achieving the improvements in DNA fragment analysis speed and throughput required by the Human Genome Project.
