TGE of all exons of all genes implicated in non-syndromic SNHL, including non-syndromic SNHL mimics, was completed as described, targeting 89 genes as part of the OtoSCOPE® v5 platform. Libraries were prepared using a modification of the solution-based Agilent SureSelect target enrichment system (Agilent Technologies, Santa Clara, CA).11 (link)In brief, 3μg gDNA was randomly fragmented to an average size of 250 bp (Covaris Acoustic Solubilizer; Covaris Inc., Woburn, MA), fragment ends were repaired, A-tails were added, and sequencing adaptors were ligated before the first amplification. Solid phase reverse immobilization purifications were performed between each enzymatic reaction. 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 prehybridization 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 (Illumina Inc, San Diego, CA) in pools of 4-6 or 48, respectively, using 100-bp paired-end reads.