GSM4432449: FM-4; Homo sapiens; miRNA-Seq
Exosome was isolated from serum using the RiboTM Exosome Isolation Reagent (Ribobio),and Exosomal RNA was extracted by the MagZol (Magen) according to the manufacturer's protocol. RNA purity was assessed using the K5500.RNA integrity was evaluated using the Agilent 2200 TapeStation (Agilent Technologies, USA) and each sample had the RINe above 7.0. RNAs were ligated with 3'RNA adapter, and followed by 5'adapter ligation. Subsequently, the adapter-ligated RNAs were subjected to RT-PCR and amplified with a low-cycle. Then the PCR products were size selected by PAGE gel according to instructions of NEBNext® Multiplex Small RNA Library Prep Set for Illumina® (Illumina, USA). The purified library products were evaluated using the Agilent 2200 TapeStation and diluted to 10 pM for cluster generation in situ on the HiSeq2500 single-end flow cell followed by sequencing (1×50 bp) on HiSeq 2500.