SBIR-STTR Award

Rapid, high-throughput DNA electrophoresis instruments are under intense scientific and commercial development at several institutions. These next-generation instruments have the potential of delivering the order-of-magnitude increases in DNA fragment analysis speed and throughput required for genomic research. However, to realize these potential benefits, advances in separation media are critically needed. We will develop a new, proprietary separation media technology with enabling features for narrow channel electrophoretic methods, such as capillary array electrophoresis. This new media technology will combine the best features of crosslinked gels and polymer solutions, i.e., high resolution and replaceability.The Phase I objectives are to(1) develop replaceable matrices for capillary electrophoresis of dsDNA with resolution equal to or greater than crosslinked polyacrylamide; and(2) demonstrate the feasibility of separating sequencing fragments with single base resolution in replaceable matrices of the type proposed here.In Phase II, the sequencing matrix will be optimized to equal or surpass the resolution and read length of crosslinked polyacrylamide.National Center for Human Genome Research (NCHGR)

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. PROPOSED COMMERCIAL APPLICATION: 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.

Thesaurus Terms:
DNA, electrophoresis, gel, method development, nucleic acid sequence