GSM5396567: PDR-serum; Homo sapiens; ncRNA-Seq
Total RNA was isolated from exosomes using the Trizol (invitrogen) according to the manufacturer instruction. RNA purity was assessed by ND-1000Nanodrop required dA260/280≥1.8, A260/A230≥2.0. RNA integrity was evaluated by Agilent 2200 TapeStation (Agilent Technologies, USA) requiring RIN≥7.0. Briefly, rRNAs were removed from total RNA using Epicentre Ribo-Zero rRNA Removal Kit (illumina, USA), then RNA was treated with RNase R (Epicentre, USA) and fragmented to approximately 200bp. Fragmented RNA (the average length was approximately 200 bp) were subjected to first strand and second strand cDNA synthesis following by adaptor ligation and enrichment with a low-cycle according to instructions of NEBNext® Ultra™ RNA Library Prep Kit for Illumina (NEB, USA). The purified library products were evaluated using the Agilent 2200 TapeStation and Qubit®2.0(Life Technologies, USA). The libraries were paired-end sequenced (PE150, Sequencing reads were 150 bp) at Guangzhou RiboBio Co., Ltd. (Guangzhou, China)using IlluminaHiSeq 3000 platform