Comparative study by 454 sequencing of small RNA content of exosome-like vesicles and total cells, data from exosomes
Exosome-like vesicles were purified from 150 mL supernatant of MonoMac-6 cells grown in RPMI 1640 supplemented with 10% FBS and OPI supplement (Sigma) by differential centrifugation . Centrifugation steps were as follows: 400 g (5 min), 1200 g (5 min), 10 000g (30 min), 100 000g (1h30). Exosome-like vesicles were resuspended in 250 uL PBS, as were cells and RNA was purified by both using 750 uL of Trizol LS. RNA from three preparations of exosomes and cells was pooled for library preparation. Two ug of exosome RNA was used and 200 ug of cell RNA for preparation of libraries.