![]() ![]() Our device thus provides a new way to generate microbeads with high throughput and no dead volume for biochemical applications. ![]() We demonstrate the potential of this method by performing on-chip a temperature-triggered DNA isothermal amplification in agarose microbeads. 3 Division of Life Science, Hong Kong University of Science and Technology, Hong Kong. 2 Natural and Environmental Sciences, Zittau/Görlitz University of Applied Sciences, Germany. The transition from agarose microdroplets to microbeads provides additional stability and facilitated manipulation. 1 Division of Life Science, Hong Kong University of Science and Technology, Hong Kong. Emulsions for PCR comprise an oil phase, a surfactant, and an aqueous phase which functions as a micro-reactor. Beads coated with primers and a DNA library are dispersed into droplets together with the second free primer. The cooling system consists of two copper wires embedded in the microfluidic device. For emulsion PCR we found that somewhat longer extension times help to ensure full-length amplicon formation and good overall efficiency of the reaction, which in turn leads to increased yield of the desired product. Schematic of emulsion PCR (ePCR) on microbeads. The agarose phase present at the inlet is thus aspirated in the device, and segmented in microdroplets. ![]() DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions bioRxiv, DOI: 10. 2020 - Multiplexed Characterization of Protein Families with DropSynth Jan. This method consists in pushing the oil continuous phase only, while suction is applied to the device outlet. DropSynth Summary from Calin Plesa on Vimeo. Alginate Microbeads are one of the most widely investigated cell encapsulation materials as they are biocompatible. We used a flow-focusing microchannel network and successfully generated agarose microdroplets at room temperature using the “push-pull” method. This emulsion PCR (ePCR) method allowed spatial isolation of each template to a single bubble and enabled clonal amplification of the template on each bead. Here, we report a new method for generating such agarose-in-oil microdroplets on a microfluidic device, with minimized inlet dead volume, on-chip cooling, and in situ monitoring of biochemical reactions within the gelified microbeads. Recently, the combination with a sol-gel switch ability, using agarose-in-oil microdroplets, has increased the range of possible applications, allowing for example the capture of amplicons in the gel phase for the preservation of monoclonality during a PCR reaction. Water-in-oil microdroplets offer microreactors for compartmentalized biochemical reactions with high throughput. ![]()
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