GSM8655802: RPE cells from 1mo rd6 mice - replicate 3; Mus musculus; RNA-Seq
To isolate RPE from the enucleated mouse eyes, anterior segment and the neural retina were removed from the eyecup, which was then incubated in RNAprotect cell reagent (Qiagen), following a previously established protocol (Xin-Zhao Wang et al. Exp Eye Res. 2012 Sep:102:1-9. doi: 10.1016/j.exer.2012.06.003. ) Total RNA was isolated from RPE pooled from both eyes per sample using RNeasy Micro kit (Qiagen) according to the manufacturer's directions and analyzed using Bioanalyzer assay (Agilent) to ensure good quality of the obtained samples (RNA integrity number, RIN ≥8). RNA samples were processed in Transcriptomics and Deep Sequencing Core (Johns Hopkins University) for library preparation and sequencing. Briefly, total RNA (~100ng per sample) was used for library preparation by following Illumina TruSeq Stranded Total RNA Library Prep Kit protocol. Libraries were then sequenced (paired-end 100 cycles) with the Illumina NextSeq 500 platform, yielding approximately 40 million raw reads per library.