Phase I demonstrated the feasibility of Akonni's gel drop microarray workflow as a complete solution for forensic SNP-typing applications. Sample preparation, PCR, and the microarray were packaged into prototype flow-through microfluidic modules for SNP discrimination. Phase II focused on refining protocols and assay chemistries, as well as packaging components and reagents into an integrated system for automated, sample-to-answer results. The system consisted of the instrument and a disposable, integrated cartridge. The instrument is composed of liquid handling, Akonni Bladder Thermal Cycler, Akonni Reader, and cartridge docking station. The integrated cartridge consists of Akonni TruTip, Akonni PCR and TruArray flow cell chambers, microfluidic circuits, and microfluidic valves. Performance criteria for the feasibility study included the capability of demonstrating a multiplex PCR and isothermal Allele-specific Arrayed-Primers Extension test for eye color; positive results from blood samples processed on the integrated system; positive results on the integrated system using lyophilized assay reagents, and correct SNP typing of mock forensic samples on the integrated system. The major product from this phase is a final technical report that supports substantial progress on these tasks. Phase III will continue defining protocols and assay chemistries, add additional SNP markers, and further refine the packaging of components and reagents in the integrated system. Plans are underway to reconfigure the platform in Phase III so as to allow processing and analyzing up to eight samples in parallel. Low cost molding of the disposable will be improved in the updated design. Developing an intuitive software and graphical user interface, as well as reducing the instrument footprint remain high priorities. 34 figures and 24 references
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