GSM4891029: Rhino-lib11: NWR iBCL2-GFP-iPSCs naïve N2B27 - rep2; Ceratotherium simum cottoni; RNA-Seq
Total RNA was was isolated using RNeasy mini kit (Qiagen, #74104) according to the manufacturer's instructions. 3 µg of isolated RNA were treated with TURBO DNase (Life Technologies, #AM2239) followed by clean up using RNeasy MinElute RNA cleanup kit (Qiagen, #74204). RNA purity and quality were assessed with microcapillary electrophoresis on Agilent 2100 Bioanalyzer (RNA Pico 6000 kit, Agilent, #5067-1513). Ribosomal RNA was depleted from 1 µg of total RNA using RiboZero Gold (Human/Mouse/Rat) kit (Illumina), followed by purification with RNeasy MinElute RNA cleanup kit. Sequencing libraries were prepared from rRNA-depleted RNA using TruSeq Stranded total RNA LT kit (Illumina, #20020598) according to the manufacturer's instructions. The final PCR amplification included 11 cycles followed by purification with Ampure XP beads (Beckman-Coulter, #A63881). Quality of final libraries was evaluated by Agilent 2100 Bioanalyzer using DNA 1000 kits (Agilent, #5067-1504). DNA concentrations were measured with Qubit (Life Technologies) and dsDNA HS Assay Kit (Thermo Fisher Scientific, #Q32854). Libraries were sequenced using Illumina NextSeq 500 instrument [75-nt single-end reads, NextSeq 500/550 High Output Kit v2.5 (75 Cycles), Illumina, #20024906].