title: GSM4459551: A12-08; Homo sapiens; miRNA-Seq
library_construction_protocol: The UC method was optimized according to the method previously described (Théry C. et al. 2006). After thawing at 37 C, plasma samples were centrifugated at 3,000 g for 15 min to remove cell debris. Then, the supernatant was diluted by seven-fold volume of PBS, centrifuged at 13,000  g for 30 min, and processed through a 0.22 μm filter to remove large particles. The supernatant was ultracentrifuged using a P50A72-986 rotor (CP100NX; Hitachi, Brea, CA, USA) at 100,000 g, 4 C for 2 h to pellet the exosomes. The pellet was resuspended in phosphate-buffered saline (PBS) and centrifuged again at 100,000 g 4 C for 2 h. After PBS washing, the exosomes pellet was re-suspended in 100 l PBS.  Total RNA was extracted and purified from plasma exosome using miRNeasy® Mini kit (Qiagen, cat. No. 217004) according to the kit instruction. RNA degradation and contamination, especially DNA contamination, was monitored on 1.5% agarose gels. RNA concentration and purity were evaluated using the NanoDrop 2000 Spectrophotometer (Thermo Fisher Scientific, Wilmington, DE) and the RNA Nano 6000 Assay Kit of the Agilent Bioanalyzer 2100 System (Agilent Technologies, CA, USA). RNA libraries were prepared for sequencing using standard Illumina protocols