title: WES for subject 260-1131, primary tumor
design: WXS library  was prepared the KAPA HyperPrep Kit with Library Amplification and pooled and underwent lyophilization using the Biotage SPE-DRY.  Post lyophilization, custom exome bait (TWIST Biosciences) along with hybridization mastermix was added to the lyophilized pool prior to resuspension. After capture and PCR amplification, library pools were quantified using qPCR (automated assay on the Agilent Bravo), using a KAPA Biosystems kit with probes specific to the ends of the adapters. Based on qPCR quantification, pools were normalized using a Hamilton Starlet to the required loading concentration. Up to 24 samples were sequenced per lane on Illumina's NovaSeq S4 sequencing technology. Cluster amplification of library pools was performed according to the manufacturer's protocol (Illumina) using Exclusion Amplification cluster chemistry and NovaSeq S4 flowcells. Flowcells were sequenced on Sequencing-by-Synthesis chemistry for NovaSeq S4 flowcells using paired 151bp runs.