Abstract 2282: Efficient sample preparation with Covaris AFA® technology enhances confidence in Illumina® TruSight™Oncology 500 Assay

Abstract High throughput and robust DNA fragmentation is a critical step in Next Generation Sequencing (NGS) for clinical genomics, ultimately ensuring higher quality for the final sequencing data. The typical requirements in NGS workflow (nucleic acid fragmentation, library preparation, sequencing, etc) comprise scalability, efficiency, reliability, and robustness. Failures to achieve confidence in any of the steps can negatively impact sequence data, rendering them unusable for diagnosis. Covaris’ Adaptive Focused Acoustics® (AFA®) technology enables efficient DNA fragmentation regardless of DNA concentration and starting molecular weight, ensuring a tight fragment size distribution, minimal sample loss, and most importantly, desired reliability and reproducibility.Illumina®’s TruSight™ Oncology 500 (TSO 500) protocol describes an enrichment-based approach to convert DNA and RNA extracted from formalin-fixed paraffin embedded (FFPE) tissue samples into libraries enriched for cancer-related genes that can be sequenced on Illumina® sequencing systems. The TSO 500 assay is optimized to prepare libraries from genomic DNA (gDNA) that is sheared to optimized fragment sizes and offers high fidelity determination of DNA variants across 523 cancer relevant genes. The TSO 500 assay is the assay of choice across most cancer centers using a standardized pipeline and therefore is suitable for routine use in precision oncology as a comprehensive genomic profiling solution. This study focuses on demonstrating the optimized DNA fragmentation protocol for TSO 500 assay with parallel 8-sample processing with an ML230 Focused-ultrasonicator. Citation Format: Debadeep Bhattacharyya, Kasturi Pal, Eugenio Daviso, Ulrich Thomann. Efficient sample preparation with Covaris AFA® technology enhances confidence in Illumina® TruSight™Oncology 500 Assay [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2282.