GSM8381176: 1744; Homo sapiens; miRNA-Seq
Blood samples were collected in EDTA tubes and centrifuged at 2500 rpm for 10 minutes. Once plasma was obtained, each sample was stored at −80°C until RNA extraction. 400 µL of plasma were used to isolate cell-free total RNA (including miRNAs) using the miRNeasy Serum/Plasma kit (Qiagen, Valencia, CA, USA) following the manufacturer's protocol. The libraries were generated and indexed using a modified Illumina TruSeq small RNA protocol. In this modified protocol the libraries were size selected (range 90-170pb) using the Blue Pippin instrument (Sage Science, Beverly,MA, USA). A positive RNA control was included (Thermo Fisher Scientific Human Brain Total RNA catalogue #AM7962). SmallRNA-sequencing (smallRNAseq) was performed by single-end sequencing on Illumina NextSeq550 platform on High Output 1x50pb RUN (NextSeq 500/ 550 High Output v2 75 cycles kit, FC-404-2005).