(1155-D) Engineering a Novel Automated DNA Library Preparation Platform from Peripheral Blood for Next-Generation Sequencing

Abstract: Next Generation Sequencing (NGS) is a powerful tool for genomic analysis and is becoming readily available, however, preparing sequencing libraries from peripheral blood is still very time-consuming and complex. Here, a fully automated system benchtop instrument can prepare sequencing libraries directly from human whole blood in a single workflow, reducing hands-on time significantly to only 20 minutes with no mid-run intervention. The instrument’s performance is evaluated for preparing sequencing libraries for whole genome sequencing (WGS) using a magnetic-based extraction process from whole blood and enzymatic fragmentation with adapter ligations process to prepare sequencing libraries. Different blood samples anticoagulated with either Sodium Citrate or EDTA were compared, and tested, along two different storage conditions, 4°C and -20°C. With an input of 40 μL of Fresh and Frozen Human Whole Blood, a 30 μL eluate of 20.6 – 37.0 ng/μL and 17.3 – 38.8 ng/μL of dsDNA were extracted, respectively for sequencing. The extracts from the automation system exhibited an average A260/A280 ratio of 1.98 across each storage condition, all comparable to a manual workflow. The automation system successfully prepared sequencing libraries from all sample types, with similar yields and molecular weight distribution libraries prepared manually on the benchtop, at an average size of 650 bp. Both the automation system and manual workflow exhibited highly similar sequencing results. Overall, the presented automation system is a reliable and user-friendly platform for automating the direct preparation of sequencing libraries from human peripheral whole blood. It is highly suitable for labs of all sizes with limited technical expertise and can help streamline the NGS workflow in multiple decentralized settings.