(1032-A) Capillary-based automation of preparing sequencing-ready libraries for nanopore sequencing technology.

Abstract: Nanopore technologies for sequencing long DNA molecules rapidly advance, leading to improved accuracy, read length, and accessibility. However, the sequencing requires a complex and time-consuming sample preparation process. Presented is a system capable of fully automating extraction, repair, end-prep, adapter ligation, and clean-up required for ligation-based library preparation from biospecimen such as peripheral blood. The system uses capillary action mixing and liquid handling to perform fast, robust, high-quality libraries from a double-stranded genomic DNA at low throughput. High molecular weight gDNA at 70kb at high purity could be extracted from human peripheral blood to generate sequencing-ready libraries. The automation system and the manual workflows yielded 2.67 ± 0.43 ng/μL (n=6) and 5.32 ± 0.22 ng/μL (n=2) of sequencing libraries, respectively. Additionally, the automated system and the manual workflows had an accuracy of ~95% and ~91% of basecalling from the same gDNA source, while the average alignment to human genome 38 was at ~96.0% and ~91.4%, respectively. Overall, the presented workflow of an automated library preparation system has the potential to revolutionize nanopore sequencing by making the process more robust and more accessible to a wider range of users.