The TGE of all exons of all genes implicated in NSHL, including NSHL mimics, was completed as described, targeting 66 or 89 genes as part of the OtoSCOPE® v4 or v5 platforms, respectively (see Supporting Information, Table S1 for full list)(5 (link)). Libraries were prepared using a modification of the solution-based Agilent SureSelect target enrichment system (Agilent Technologies, Santa Clara, CA)(6 (link)). In brief, 3μg of gDNA was randomly fragmented to an average size of 250 bp (Covaris Acoustic Solubilizer; Covaris Inc., Woburn, MA), and adaptor ligated before the first amplification. Hybridization and capture with RNA baits was followed by a second amplification before pooling for sequencing. Minimal amplification was used – typically 8 cycles for the pre-hybridization PCR (range 8–10 cycles) using NEB Phusion HF Master Mix (New England BioLabs Inc, Ipswich, MA) and 14 cycles for the post-hybridization PCR (range 12–16 cycles) using Agilent Herculase II Fusion DNA Polymerase. All samples were barcoded and multiplexed before sequencing on either an Illumina MiSeq or HiSeq system (Illumina Inc, San Diego, CA) in pools of 4–6 or 48, respectively, using 100-bp paired-end reads.