GSM8244939: OPA1+/+ cornea, 3 months, replicate #2; Mus musculus; RNA-Seq
Total RNA was extracted usingTrizol reagent and purified afterwards with Qiagen RNeasy kit following the manufacturer's procedure. Total RNA concentrations were measured with Qubit and quality was assessed using RNA ScreenTape Assay (Agilent TapeStation 4200). 500 ng of good quality (RIN >8.8) total RNA was ribo-depleted using Illumina's mammalian Ribo-Zero Magnetic Gold Kit HMN/Mouse/Rat (MRZG12324). Library preparation was completed on ribo-depleted RNA using the NEBNext Ultra Directional RNA Library prep kit (NEB, #E7420L) using 8 cycles of PCR amplification and indexed using single (i7) indexing. Indexed library preps from each sample were then pooled and sequenced with 75SE reads at a pool concentration of 1.4 pM on the NextSeq 500 using a NextSeq High Output 75 cycle flow cell (Illumina).