Abstract 259: Fully automated targeted sequencing sample prep on open platform liquid handlers

Abstract

Abstract To accommodate the growing demands of sample throughput, reliability, and reduced hands-on time of targeted sequencing, automation is critical. The implementation of sample preparation automation is limited by several factors — complicated protocols and long incubation times. As an industry first, Avida’s Point-n-Seq™ hybridization-based targeted sequencing kit solution features 1 hr hybridization time and PCR free capture which enables full automation on a broad range of open platform liquid handlers, from the economical Opentrons (OT-2) to the high throughput Hamilton (Microlab Star). We report a walk-away targeted sequencing sample-prep solution based on the highly efficient chemistry of our Point-n-Seq technology. This strategy supports PCR-free capture for variant detection. A sequencing-ready captured library is prepared in 6 hours, with just 25 minutes of hands-on time. More importantly, a PCR-free protocol increases sequence uniformity, decreases amplicon bias as well as GC-bias, and eliminates the need for purification and quantification before capture. We have demonstrated the feasibility of automated workflows using 40 ng of genomic DNA with panel sizes ranging from 20 Kb to 2 Mb. Using a comprehensive oncology panel containing 500 genes to analyze Horizon oncology cfDNA (30 ng) or FFPE gDNA (40 ng) reference standards, we demonstrated reliable detection of SNV and structural variations with allele frequency matching the expected values. The Point-n-Seq technology enables fully automated targeted sequencing sample prep on open platform liquid handlers, enhancing accuracy and reliability while reducing hands-on time. The one hour hybridization time makes hybridization more economical on the liquid handler when compared to off-instrument for overnight/longer hybridization. This automated solution greatly accelerates the development of fast, simple, and accurate genomic profiling assays. Furthermore, Point-n-Seq targeted methylation sequencing and targeted RNA assays can also be easily adapted to the same automation platforms for research and clinical applications. Citation Format: Ashraf Wahba, Yun Bao, Heng Wang, Kinnari Watson, Grace Q. Zhao, Shengrong Lin. Fully automated targeted sequencing sample prep on open platform liquid handlers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 259.