Microfluidic Sample Preparation Device for Environmental DNA Separation and Washing **********

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Detection of environmental DNA (eDNA), excreted from organisms into the environment at very low concentrations, has emerged as a powerful method for identification of dangerous or at-risk species in aquatic systems. Precision Biomonitoring (PBI) offers a ground-breaking technology for detection of rare eDNA on-field. It includes a sample preparation unit for filtration of freshwater samples and a detection unit for sensing eDNA. Company's problem is associated with the complex freshwater properties making sample filtration complicated, while the suspended matter in the samples act as inhibitors negatively affecting the detection limit and sensitivity of the system. The above problem is significant for which a sample preparation solution must be researched that is 1) effective on a broad range of water samples, 2) integrate-able with company's platform, 3) portable and battery-operated, and 4) low cost.**We propose developing a microfluidic particle separation, DNA isolation and DNA washing device to meet the attributes of environmental sample preparation. Multi-radius serial curved microchannels will be used to sequentially extract inhibitory particles from the sample using elastic and inertial forces that can be applied to particles of interest based on their sizes. Two Research Assistants (RAs) and one PhD student will conduct a two-phase research. In Phase 1, we will characterize the particle and biological content of water samples. In Phase 2, we will use the information in Phase 1 and our knowledge of fluid-particle flow to develop and test the microfluidic device. Particles from large to small will be sequentially separated from the sample and the DNA content, captured on microbeads, will be separated from the other molecular content and washed into clean buffers for detection with the PBI system. Our proposed device is innovative because it will be capable of purifying DNA molecules that are already dispersed at very low concentrations among the inhibitory particles, while being able to process complex fluids with non-Newtonian properties instead of the commonly-used water-based solutions. **