Automated, high throughput, microfluidic assembly and transfer ofsynthetic genomes (SynGen system)

Synthetic biology represents a potentially critical biotechnological solution for therapeutics development, materials production, diagnostics, and manufacturing capabilities and paradigms. But limited ability to assemble, transfer, and transfect large DNA constructs remains a fundamental barrier. To address this urgent DARPA need, ChromoLogic LLC proposes to develop the SynGen system, a microfluidic platform for the automated, pipet-free, step-wise assembly of commercial synthetic error-free DNA building blocks (~4-10kbp) into large DNA constructs (>100kpb). The SynGen system is constituted of the G-Cart disposable cartridge composed of micro-channels and incubation chambers to gently transfer, assemble (using the Gibson method), and prepare large DNA construct for transfection; the DNAflow platform composed of micro-pump, valves and heating pad to precisely regulate microfluid movements and temperature in the inserted G-Cart; and the SoftGen customized software that controls the DNAflow device for the implementation of procedures. In less than 3 hours, the compacted, purified, long DNA construct is collected into a sterile tube, ready for transfection into the microorganism of interest using standard protocols and with a transfection efficiency of = 1%. The synGen system provide a high-fidelity, low recombination rate, rapid, versatile, easy-to-use and low cost technology for the assembly, transfer and transfection of synthetic DNA constructs.