Electroosmotic microfluidic oligonucleotide synthesis reactor

摘要

The principle objective of this dissertation is the development of a microfluidic oligonucleotide (oligo) synthesis reactor that sequences oligos with equal quality to commercial synthesizers and with equal waste reduction to previously published microfluidic synthesizers. The microfluidic approach to oligo synthesis increases throughput due to parallelization, provides simplification by integration of multiple functions into one platform, and decreases cost due to miniaturization and waste reduction. Microfluidic oligo synthesizers demonstrate these advantages over industrial synthesizers but have error rates that are too high for commercialization.;The microfluidic synthesizer design investigated in this dissertation incorporates several innovations to reduce the error rates of previous microfluidic synthesizers. The design eliminates the need for microvalves by utilization of electroosmotic flow (EOF). The device is fabricated in photosensitive glass which is compatible with standard oligo chemistry, and facilities pmol scale reactors and embedded optics. The surface properties of a photosensitive glass are characterized to inform device design. A special coating is developed that supports EOF and prevents side reactions from occurring on channels etched into the glass. Unique microlenses are fabricated to enhance the performance of embedded optics. Custom electronics are built to control EOF and monitor electric current in the device for flow rate determination. Finally, the device is modeled in COMSOL multiphysics. The electroosmotic microfluidic oligo synthesizer has been fabricated and is currently being evaluated for oligo synthesis. Future work will expand the design for parallel synthesis of 16 unique oligos on one device.