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Spatially-separated Combinatorial DNA Assembly Device


Scientists at the JBEI are developing a device that can be used to efficiently assemble DNA parts, such as genes encoding enzymes, into multiple combinations, and then screen the resulting combinatorial library to identify combinations with the most desirable properties. The device combines into one microfluidic chip all of the steps necessary for this process: assembly of DNA parts; transformation and expression of these assemblies in whole-cells or cell-free platforms; cell culture; and functional assays using techniques such as colorimetric reporters, cell labeling-sorting, fluorescence imaging, and-or spectroscopy. The JBEI invention could be used in engineering plants and enzymes for better biofuel production, or developing crops that are more resistant to pathogens or drought. The device places each component in the processDNA parts, reagents, cells, assayable markersinto discrete droplets that flow through microfluidic channels on a chip. Specific droplets are fused at designated times and locations in the channels to precisely control every reaction and incubation step. Throughout the process, each combination of biological parts is kept spatially separated from the other combinations. Thus, each droplet comes off the chip with its function assessed and its combination of parts known. In addition, droplets can be removed from the chip at different points throughout the process to obtain various intermediate products such as recombinant DNA, transformed cells, labeled cells, or protein cocktails.
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