GSM7750345: RA_A5; Homo sapiens; RNA-Seq
Fresh peripheral blood samples were collected in EDTA-K2 anticoagulant tubes separately, transported in a 4°C incubator, and disposed of within 2 hours. PBMCs were isolated from fresh blood by density gradient centrifugation over HISTOPAQUE-1077 (Sigma-Aldrich), according to the manufacturer's instructions. Total RNA was extracted using the mirVana miRNA Isolation Kit (Ambion) following the manufacturer's protocol. RNA integrity was evaluated using the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). The samples with RNA Integrity Number (RIN) ≥ 7 were subjected to the subsequent analysis. The libraries were constructed using TruSeq Stranded mRNA LTSample Prep Kit (Illumina, San Diego, CA, USA) according to the manufacturer's instructions. These libraries were sequenced on the Illumina sequencing platform (HiSeq X Ten) and 125bp/150bp paired-end reads were generated.