Micro-Solid Phase Extraction of DNA on Microchips

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DESCRIPTION (provided by applicant): Despite the current fad that the 'post-genome' era is coming to an end, it is likely that genetic analysis will continue to be a prolific field of exploration. Genetic analytical methods will be critical in fleshing out new mutations correlative of neoplastic growth, understanding the significance of single nucleotide polymorphisms, and developing better genotyping methods for microsatellite DNA. Accordingly, the purification of nucleic acid material from crude biological samples, a requisite protocol for successful amplification of target DNA sequences by the polymerase chain reaction (PCR), is still of vital importance. Development of a protocol for solid phase extraction (SPE) of DNA from biological samples (whole blood, cell culture, etc.) has been shown to be feasible in a microchip format during the previous funding period. This platform exploits the power of microchip technology for reducing sample volumes and increasing the speed of the extraction process, but optimization is needed to maximize the potential of this method. The additional benefit of the microchip platform is the ability to integrate additional processing steps. This will be investigated through the development of a microchip SPE-based DNA purification protocol that is directly and seamlessly mated with on-chip PCR, setting the stage for truly integrated genetic analysis on a microchip (a.k.a., a true lab-on-a-chip). A major part of the development work will also focus on integration of DNA quantitation methods directly on the microchip between the SPE and PCR steps. This will allow optimal use of the DNA extracted from the sample in the PCR amplification.