GSM2493745: P42_Heart_Ribo_rep1; Mus musculus; RNA-Seq
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. Sequencing libraries of ribosome protected fragments were generated using ARTseqTM Ribosome Profiling Kit (Epicentre, RPHMR12126) according to the manufacturer's instructions. And, sequencing libraries of poly(A)+ RNA were  generated using VAHTSTM mRNA-seq v2 Library Prep Kit for Illumina (Vazyme Biotech, NR601-01) according to the manufacturer's instructions.