GSM4860427: plasma small RNA-Seq VOVP102; Homo sapiens; miRNA-Seq
Total RNA from plasma samples was extracted with the miRNeasy plasma/serum mini kit (Qiagen) using the QiaCube extractor (Qiagen) following the manufacturer's instructions. RNA samples were stored until analysis at -80°C. Small RNA transcripts were converted into barcoded cDNA libraries. Library preparation was performed with the NEBNext Multiplex Small RNA Library Prep Set for Illumina (New England BioLabs Inc., USA). Multiplex adaptor ligations, reverse transcription primer hybridization, reverse transcription reaction, and PCR amplification were performed according to the protocol for library preparation (Protocol E7330, New England BioLabs Inc., Ipswich, MA, USA). After PCR amplification, the cDNA constructs were purified with the QIAQuick PCR Purification Kit (Qiagen, Germany) following the modifications suggested by the NEBNext Multiplex Small RNA Library Prep Protocol and loaded on the Bioanalyzer 2100 (Agilent Technologies, Milan, Italy) using the DNA High Sensitivity Kit (Agilent, Germany) according to the manufacturer's protocol.