GSM4504262: sba3888_n_mir; Rattus norvegicus; miRNA-Seq
A small piece of frozen aorta (≤100 mg) was cut in the Petri dish on dry ice to prevent melting. The piece of aorta was homogenized in 1 mL of mirVana miRNA lysis/binding buffer (ThermoFisher Scientific, Waltham, MA, USA) in a 5 mL tube for 1 min at maximum speed with a Polytron PT 1600 E homogenizer (Brinkmann Instruments, Mississauga, ON, Canada) equipped with a dispersing aggregate PT-DA 1607/2EC. The homogenate was centrifuged at 1500 x g for 2 min at 4 °C to remove foam generated during homogenization, and processed for total RNA extraction using the mirVana miRNA isolation kit in accordance with the manufacturer's instructions, followed by genomic DNA removal with the TURBO DNA-free kit (ThermoFisher Scientific) according to the manufacturer's protocols. The concentration and purity of the RNA were assessed with a NanoDrop ND-100 spectrophotometer (ThermoFisher Scientific). RNA quality was assessed with an Agilent 2100 bioanalyzer (Agilent Technologies, Santa Clara, CA). Samples with RNA integrity number (RIN) of ≥8 were used for RNA library construction. One microgram of total RNA (n = 6 per group) was used for small RNA library construction using the TruSeq small RNA sample preparation kit (Illumina, San Diego, CA) according to the manufacturer's protocol. Small RNAs were sequenced using the HiSeq 2500 system (Illumina, 1 x 51 bp single-end, rapid mode) with the goal to obtain 30 million reads per sample.
GEO Accession
GSM4504262