(1235-D) Automating the NEXTFLEX® Human Whole Exome Sequencing Panel Target Enrichment workflow on the Revvity Fontus NGS workstation

Abstract: Whole exome sequencing is a widely used next-generation sequencing (NGS) method to sequence the protein-coding regions of genes reducing the need to sequence the entire genome. An efficient workflow from library preparation to target capture is essential to producing reliable libraries.
Despite the advantages of hybridization capture, such as reduced bias and consistent data across diverse sample types, its intricate workflow and extended preparation time have posed challenges for users. The following discusses the automated preparation of libraries with the NEXTFLEX® Human Whole Exome Sequencing Panel (WES) on the Revvity Fontus NGS. The combination of automation and the streamlined protocol of this panel allows for cost-effective and time-saving sequencing of the whole exome.
The NEXTFLEX® WES Panel has a size of 37.1 Mb and includes the target regions of all major whole exome panels on the market. With its robust probe design and synthesis, the panel offers high coverage, uniformity, and on-target ratio even in GC-, AT-rich regions and homologous regions where other panels struggle.
Utilizing the Chemagic™ 360 instrument and chemistry, DNA of high yield was extracted from whole blood samples. The DNA samples were then prepared through the automated workflow on the Revvity Fontus NGS workstation, producing libraries comparable to manually prepared libraries.
The automated workflow offers a high throughput solution for users to process up to 96 samples per setup, includes a simple user interface including selection of sample number, desired fragmentation size, and number of PCR cycles, tailored for walk-up use. A detailed graphical user interface and positioning charts walk the user through the deck setup process, reducing the chance for human errors and overall hands-on time.
For research use only. Not for use in diagnostic procedures.