GSM9063446: AG-ZT6-4; Mus musculus; RNA-Seq
At each time point, four mice from each group were euthanized by cervical dislocation, and the right eyes were enucleated. Under dim red light, the anterior segments were removed, and the retinas were carefully dissected. Total RNA was isolated from retina tissues using TRIzol reagent according to the manufacturer's instructions. RNA integrity was assessed using the Nano 6000 Assay Kit on the Bioanalyzer 2100 system. mRNA was enriched from total RNA using poly-T oligo-attached magnetic beads. Fragmentation was performed using divalent cations at elevated temperatures in First Strand Synthesis Reaction buffer. Approximately 2 μg of total RNA was used to synthesize first-strand cDNA using random hexamer primers, and second-strand cDNA synthesis was subsequently conducted with DNA polymerase I and RNase H. After end-repair of DNA fragments, adaptors with hairpin-loop structures were ligated for hybridization. The cDNA fragments, preferentially 370-420 bp in length, were PCR-amplified with Phusion High-Fidelity DNA polymerase and universal PCR primers.