This Phase II project continues the development and application of new materials, especially magnetic microparticles for directly isolating DNA from cells, tissues, and biological fluids, including whole blood, in an extremely rapid format. The materials eliminate the need to perform a lysis step and capture DNA from complex materials thereby simplifying sample preparation methods. The functionalized microparticles for binding and releasing nucleic acids utilize a proprietary chemistry which enables them to spontaneously and rapidly lyse cells and capture the exposed DNA in situ under virtually any biological conditions. As a result DNA can be isolated from whole blood without any lysis step. The unique surface phosphonium group chemistry causes nucleic acids to bind with exceptional strength until released by a unique cleavage mechanism where a linker group is selectively broken by a simple chemical reaction in order to free DNA from the solid surface. Release by chemical cleavage rather than traditional elution prevents premature, uncontrolled release of material and yields pure nucleic acid. Isolated DNA is compatible with downstream applications such as PCR. Additional linking groups and cleavage methods are being explored. Phase II will develop the expertise to perform the complete synthesis including preparing the magnetic cores, coating the cores, and functionalizing with linker and head groups. The capability to manufacture the fuctionalized magnetic particles on a large scale will be developed. We have produced a prototype kit for isolating DNA from whole blood. We areing the range of applications of these new materials is being expanded to plasmid purification, isolation of DNA from viruses, and capture of DNA from infectious microorganisms in urine, foodstuffs and culture media with no further isolation steps. The materials are expected to streamline sample preparation and DNA extraction across a broad variety of applications. This Phase II project continues the development and application of new materials, especially magnetic microparticles for directly isolating DNA from cells, tissues, and biological fluids, including whole blood, in an extremely rapid format. The materials eliminate the need to perform a lysis step and capture DNA from complex matrices thereby simplifying sample preparation methods. The materials will find use in a great number of applications in the life sciences and diagnostics industries for which DNA extraction is necessary. ? ? ?
