GSM5289810: pooled RNA from H09-69 and C81 [0A]; Homo sapiens; miRNA-Seq
Fresh venous blood 20 ml was collected from each participant on awakening, and immediately transferred to a tube containing 3.2% sodium citrate (1:10 dilution). PBMCs were isolated by Ficoll-Hypaque gradient centrifugation (HISTOPAQUE®-119, Sigma-Aldrich, Inc.,USA), and then stored in RNAlater (Ambion Inc., USA) at -80°C until RNA isolation. A miRNeasy Mini Kit column-based system (Qiagen, Valencia, CA, USA) was used for isolation of total RNA, and treated with DNase according to the manufacture protocol. RNA samples were run on a RNA 6000 Nano Gel System (Agilent Technologies Inc., CA, USA) using an Agilent 2100 Bioanalyzer (Agilent) to determine the quality of RNA. Only samples with A260/A280 ratios of 1.9 to 2.1 and RNA Integrity Number >= 8 were used for further analysis. The pooled RNA libraries were subject to NGS assay to determine miRNA expression profile. Illumina MiSeq platform was adopted for the profiling job. The RNA libraries were first prepared in accordance with TruSeq Small RNA Sample Preparation protocol (Illumina) followed by sequencing with MiSeq platform.