Abstract 4087: Panel-adjustable and user-friendly hybridization-capture technology delivers high performance demonstrated by different exome probe panels

Targeted sequencing by hybridization-capture is a versatile and powerful way to interrogate selected regions of interest in large and complex genomes. However, the performance of a given hybridization-capture workflow is often optimized only towards a specific approach to probe panel design. Here we demonstrate a hybridization-capture enrichment system that delivers high performance on three commercially available Exome panels with short and overnight hybridizations. This is achieved using our new xGen Hybridization and Wash v3 (xGen v3), which features a simple user-friendly workflow without any pre-heating, hot-liquid handling, or time-and-temperature sensitive steps. Human genomic reference libraries were constructed with xGen™ cfDNA & FFPE DNA Library Preparation Kit using Coriell NA12878 gDNA. These equivalent libraries are hybridized to xGen™ Exome Hyb Panel v2, xGen™ Exome Hyb Panel v1, or a double-stranded-probe Exome panel. Hybridization incubations from 1-hour to overnight were compared, spanning total inputs from 100ng to microgram quantities per hybridization-capture. xGen v3 delivered consistent performance for all three Exome panels, without the increase in GC-bias that typically occurs for short hybridizations. Benchmarking using a currently available commercial kit is presented for performance comparison. Non-biased sampling reveals properties of the library as directed by the panel design. The demonstration here displays a panel-directed enrichment workflow that is reliable for wide range of total capture inputs and hybridization times for each given Exome panel, spanning different panel design approaches as well as single- and double-stranded probes. This behavior can be extrapolated to applications that requires multiplexing, low-pass or deep-sequencing needs, and custom panels. The panel-directed consistency over a large range of inputs and hybridization times makes xGen v3 adaptable to a large variety of applications, from quick surveys to high-throughput and/or in-depth interrogations as directed by the panel design. Citation Format: Alexandra Wang, Daniel Saatman, Ekaterina Star, Jinglie Zhou. Panel-adjustable and user-friendly hybridization-capture technology delivers high performance demonstrated by different exome probe panels [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 4087.