DNA technologies from PCR to allele discrimination are now common and indispensable techniques utilized in a myriad of fields such as healthcare and agriculture. While traditional bench top methodologies are well refined, the standard techniques often require sample volumes and costs that are prohibitive for high throughput applications. Thus genetic screening at rapid rates and low costs is a requirement for further propagation of DNA technologies in large scale operations. Microfluidic platforms are particular well-suited to meet the challenge of creating technologies capable of high throughput, continuous flow, multiplexed allelic discrimination. Specifically, by reducing a typical reaction system from a milliliter scale Eppendorf tube down to nanoliter sized droplets, genetic screening may be performed at a fraction the cost. Here, we present two novel poly(dimethylsiloxane) (PDMS) microfluidic platforms capable of multiplexing single nucleotide polymorphism (SNP) detection. Utilizing unique SNP assays (Invader and Taqman PCR) well suited for microfluidics applications, both platforms include onchip optical detection of fluorophores that allow for direct allelic read out. Utilizing benchtop amplified target DNA, successful SNP detection on-chip was achieved in the first device with unambiguous signal readout spanning nearly 80 target DNA/probe combinations. In the second device, both target DNA amplification and allele detection were performed on-chip. Taken together, our novel PDMS microfluidic platforms provide a key advance in microfluidic devices for allele discrimination. Device capableof high throughput and affordable genomic screening now looms.
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CONTINUOUS FLOW, HIGH-THROUGHPUT MICROFLUIDIC PLATFORMS FOR GENOMIC DISCRIMINATION ASSAYS
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