NextSeq 500 paired end sequencing; RNA-seq of E18.5, C57BL6 mouse retinal cells treated with glucose starvation compared with untreated controls
Retinas were isolated from E18.5, C57BL6 mouse embryos and digested with 0.05% trypsin (Gibco) for 15 minutes at 37 C.  After 1 day of culture, cells were starved for glucose in Neobasal medium supplemented with B27 supplement without insulin, 0.06 g/L glutamine, 1% penicillin-streptomycin (Gibco) and 1 mM HEPES for 6 h. The cells were harvested for RNA isolation after additional 6 days of culturing. Normal cultured cells were used as control.  RNA extracts were isolated by using TruSeq Stranded Total RNA with RiboZero.  Reverse transcription was done by using RevertAid first strand cDNA synthesis kit (ThermoFisher)