title: Automated digital microfluidic sample preparation for next-generation DNA sequencing.
abstract: Next-generation sequencing (NGS) technology is a promising tool for identifying and characterizing unknown pathogens, but its usefulness in time-critical biodefense and public health applications is currently limited by the lack of fast, efficient, and reliable automated DNA sample preparation methods. To address this limitation, we are developing a digital microfluidic (DMF) platform to function as a fluid distribution hub, enabling the integration of multiple subsystem modules into an automated NGS library sample preparation system. A novel capillary interface enables highly repeatable transfer of liquid between the DMF device and the external fluidic modules, allowing both continuous-flow and droplet-based sample manipulations to be performed in one integrated system. Here, we highlight the utility of the DMF hub platform and capillary interface for automating two key operations in the NGS sample preparation workflow. Using an in-line contactless conductivity detector in conjunction with the capillary interface, we demonstrate closed-loop automated fraction collection of target analytes from a continuous-flow sample stream into droplets on the DMF device. Buffer exchange and sample cleanup, the most repeated steps in NGS library preparation, are also demonstrated on the DMF platform using a magnetic bead assay and achieving an average DNA recovery efficiency of 80%±4.8%.
authors.affiliation: Sandia National Laboratories, Livermore, CA 94550, USA.
authors.forename: Hanyoup
authors.initials: H
authors.lastname: Kim
authors.forename: Michael S
authors.initials: MS
authors.lastname: Bartsch
authors.forename: Ronald F
authors.initials: RF
authors.lastname: Renzi
authors.forename: Jim
authors.initials: J
authors.lastname: He
authors.forename: James L
authors.initials: JL
authors.lastname: Van de Vreugde
authors.forename: Mark R
authors.initials: MR
authors.lastname: Claudnic
authors.forename: Kamlesh D
authors.initials: KD
authors.lastname: Patel
mesh_terms: D057205:Automation, Laboratory; D003217:Conductometry; D004247:DNA / Q000032:analysis*; D059014:High-Throughput Nucleotide Sequencing / Q000295:instrumentation / Q000379:methods; D006801:Humans; D007239:Infections / Q000175:diagnosis / Q000235:genetics*; D046210:Microfluidic Analytical Techniques; D015203:Reproducibility of Results
chemical_list: D004247:DNA