title: RNAseq of organ of Corti RNA from mice carrying human mutations in Mir96
design: RNAseq of organ of Corti RNA from mice carrying human mutations in Mir96
library_construction_protocol: 4-day old pups from heterozygote x heterozygote matings were collected and culled by decapitation. The head was bisected and the brain removed. Half heads were put in RNAlater and the organs of Corti dissected out, separated from modiolus and lateral wall, and put in fresh RNAlater on ice. Total RNA was extracted using the SPLIT RNA extraction kit (Lexogen) according to the manufacturer's instructions. Library generation was carried out according to the “TruSeq Stranded mRNA Sample Preparation Guide” with the corresponding kit [Illumina Inc. Cat.# RS-122-2101 or RS-122-2102]. Starting from 250ng of total RNA, mRNA was purified, fragmented, and primed for cDNA synthesis. cDNA first strand was synthesized with SuperScript-II Reverse Transcriptase (Thermo Fisher Scientific, Cat.#18064-014), followed by cDNA second strand synthesis using Illumina reagents, followed by A-tailing and adaptor ligation. Finally, the libraries were enriched by PCR.