GSM6997634: Plasma, SEC, rep2; Homo sapiens; miRNA-Seq
RNA was extracted from 250 µl of the plasma sample or from 250 µl EVs isolation samples using a miRNeasy Mini kit (QIAGEN) following the manufacturer's instructions. Purified RNA was eluted in 20 μl of nuclease-free water. To monitor the purification and amplification of miRNAs, 52 spike-ins (synthetic miRNAs at different concentrations) (QIAseq miRNA Library QC PCR Assay Kit) were added to the initial sample before RNA extraction. Quantity and quality of the purified RNA were determined using NanoDrop and Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA, USA). The RNA concentrations obtained ranged between 164 pg/µl and 349 pg/µl. NEBNext Multiplex Small RNA Library Prep Set for Illumina (New England BioLabs, Ipswich, MA, USA), NEXTFlex Small RNA-Seq Kit v3 (PerkinElmer, Waltham, MA, USA), and SMARTer smRNA-seq kit (Clontech Laboratories, CA, USA)