Specifically, in one application we have developed a method for high-throughput single cell DNA purification and detection of the translocation, an approach for investigating the frequency of rare mutation events within individuals prior to the onset of clinical symptoms. The subsequent analysis of the individual beads by flow cytometry, capillary electrophoresis or sequencing techniques then reveals the target presence, size or sequence, as desired. This challenge has been addressed by encapsulating cells together with primer functionalized beads in nanoliter agarose droplets which can then be used as porous carriers for sequentially performing cell lysis and cleanup followed by microemulsion PCR. A significant challenge is the release of genomic DNA from single cells in these nanoliter droplets. To enable this new analysis technology, we have developed microfluidic droplet generators that produce highly uniform 1-5 nL droplets at very high production rates that are subsequently used to perform PCR and RT-PCR analyses. In forensics, the ability to type individual cells and to analyze cellular mixtures cell-by-cell offers unprecedented identification capabilities. In many cancers, small subpopulations of distinct cell types cause or drive the disease but are difficult to study using conventional biomolecular methods. Single cell genetic analysis using nanoliter emulsion droplet technology offers the ultimate in digital sensitivity and an unprecedented view of the characteristics of cell populations that are masked by the ensemble average. “SINGLE CELL GENETIC AND FORENSIC ANALYSIS USING NANOLITER DROPLET MICROFLUIDICS”
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