Library Construction: An aliquot of genomic DNA (100-150ng in 50µL) is used as the input into DNA fragmentation (aka shearing).  Shearing is performed acoustically using a Covaris focused-ultrasonicator, targeting 150bp fragments.   Library preparation is performed using a commercially available kit provided by KAPA Biosystems (KAPA HyperPrep Kit with Library Amplification product KK8504) and IDT's duplex UMI adapters. Unique 8-base dual index sequences embedded within the p5 and p7 primers (purchased from IDT) are added during PCR. Enzymatic clean-ups are performed using Beckman Coultier AMPure XP beads with elution volumes reduced to 30µL to maximize library concentration.  Following library construction, library quantification is performed using the Invitrogen Quant-It broad range dsDNA quantification assay kit (Thermo Scientific Catalog: Q33130) with a 1:200 PicoGreen dilution. Following quantification, each library is normalized to a concentration of 35 ng/µL, using Tris-HCl, 10mM, pH 8.0. All steps performed during the library construction process and library quantification process are performed on the Agilent Bravo liquid handling system. In-solution hybrid selection: After library construction, hybridization and capture are performed using the relevant components of IDT's XGen hybridization and wash kit and following the manufacturer's suggested protocol, with several exceptions. A set of 12-plex pre-hybridization pools are created.  These pre-hybridization pools are created by equivolume pooling of the normalized libraries, Human Cot-1 and IDT XGen blocking oligos. The pre-hybridization pools undergo lyophilization using the Biotage SPE-DRY.  Post lyophilization, custom exome bait (TWIST Biosciences) along with hybridization mastermix is added to the lyophilized pool prior to resuspension.  Samples are incubated overnight.  Library normalization and hybridization setup are performed on a Hamilton Starlet liquid handling platform, while target capture is performed on the Agilent Bravo automated platform. Post capture, a PCR is performed to amplified the capture material. Preparation of libraries for cluster amplification and sequencing: After post-capture enrichment, library pools are quantified using qPCR (automated assay on the Agilent Bravo), using a kit purchased from KAPA Biosystems with probes specific to the ends of the adapters. Based on qPCR quantification, pools are normalized using a Hamilton Starlet to the required loading concentration. Up to 24 samples are sequenced per lane on Illumina's NovaSeq S4 sequencing technology. Cluster amplification and sequencing: 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.