title: GSM3672240: Plasma_10; Homo sapiens; RNA-Seq
library_construction_protocol: Exosomes were isolated from plasma using the ExoQuick exosome precipitation solution according to the manufacturer's instructions with minor modifications. The mixture was then further centrifuged at 1500 g for 30 min. The exosome pellet was dissolved in nuclease free water; RNA was extracted immediately from the solution. Library preparation was performed with the NEBNext Multiplex Small RNA Library Prep Set for Illumina (New England BioLabs Inc., USA). For each library, 6 μL of RNA (min 35 ng) were used in all the experimental procedures as starting material. Each library was prepared with a unique indexed primer so that the libraries could all be pooled into one sequencing lane. 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., 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, Germany) using the DNA High Sensitivity Kit (Agilent, Germany) according to the manufacturer's protocol. Libraries were pooled together (24plex) and further purified with a gel size selection.
attributes.tag: GEO Accession
attributes.value: GSM3672240