GSM5208756: C57Bl/6J + GSK2879552 replicate 2 [G764.04_B6_GSK]; Mus musculus; RNA-Seq
Right eye retina from each animal was flash frozen in liquid nitrogen and stored at -80oC. RNA extraction and purification was done with RNeasy Mini Kit and RNA shredder (Qiagen). RNA integrity number (RIN) was measured using BioAnalyzer (Agilent Technologies) RNA 6000 Nano Kit to confirm RIN above 7 for each sample. The cDNA libraries were prepared using the Illumina® Stranded mRNA Prep, Ligation kit (Illumina) as per the manufacturer's instructions. Briefly, polyA RNA was purified from 200 ng of total RNA using oligo (dT) beads. The extracted mRNA fraction was subjected to fragmentation, reverse transcription, end repair, 3'– end adenylation, and adaptor ligation, followed by PCR amplification and SPRI bead purification (Beckman Coulter). The unique dual index sequences (IDT® for Illumina® RNA UD Indexes Set A, Ligation, Illumina) were incorporated in the adaptors for multiplexed high-throughput sequencing.