GSM9107422: Intermittent hypoxia - ZT0 - REP1; Mus musculus; RNA-Seq
50–100 mg frozen liver was homogenised in 1 mL TRIzol Reagent (Invitrogen 15596026). RNA was isolated by chloroform extraction and isopropanol precipitation, washed in 75 % ethanol and resuspended in DEPC-treated water. Integrity was checked on Agilent TapeStation RNA ScreenTape. 1 µg total RNA → Illumina Stranded mRNA Prep, Ligation (poly-A selection). mRNA fragmented, first-strand cDNA synthesis with random primers, second-strand synthesis incorporating dUTP for strand specificity, A-tailing, adapter ligation, 10–11 PCR cycles, validation on TapeStation and Quantifluor. Libraries pooled equimolarly and sequenced 2 × 100 bp on NovaSeq 6000 (S4 XP).