Instrument: Illumina HiSeq 2500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Frozen tissue samples were lysed separately using 1 ml of mammalian lysis buffer (200 µl 5x Mammalian Polysome Buffer, 100 µl 10% Triton X-100, 10 µl DTT(100 mM), 10 µl DNase I (1U/µl), 2 µl Cycloheximide (50 mg/ml), 10 µl 10% NP-40 and 668 µl Nuclease-Free Water). After incubation for 20 min on ice, lysates were cleared by centrifugation at 10000 x g for 3 min at 4°C. Ribosomal RNA was depleted using the Ribo-Zero magnetic kit (Epicentre). Sequencing libraries of ribosome protected fragments were generated using ARTseqTM Ribosome Profiling Kit according to the manufacturer's instructions. Sequencing libraries of poly(A)+ RNA were generated using VAHTSTM mRNA-seq v2 Library Prep Kit for Illumina according to the manufacturer's instructions.