GSM8080252: cochlear and vestibular HCs, DMDO59 WT; Mus musculus; RNA-Seq
RNA was extracted from sorted cochlear and vestibular HCs using the Direct-zol RNA MiniPrep Kit (Zymo Research). First, TRIzolTM LS (InvitrogenTM) was added to each sample of HCs to 70% of total volume. All following steps were conducted as outlined in the Direct-zol RNA MiniPrep Kit protocol, eluting RNA in 25µL of nuclease free water in the final step. Libraries were prepared from 2.75-5.43ng of RNA from each sample using the NEBNext Ultra Directional RNA Kit per manufacturer's instructions with a total of 22 PCR cycles, except for a 5-minute fragmentation time instead of 15 minutes and a SPRI bead size selection before PCR.