GSM4610718: NC-2; Homo sapiens; RNA-Seq
Blood samples were centrifuged at 1,500 g for 15 minutes at 4 °C. The collected supernatant (2.5 mL plasma) were then centrifuged at 12,000 g for 30 minutes at 4 °C. The supernatant was collected and ultra-centrifuged at 100,000 g for 2 hours at 4 °C to pellet the exosomes. Exosome pellets were then washed in filtered PBS and re-centrifuged at 100,000 g, the supernatant was removed and the final exosomal pellet was re-suspended in 200 µl PBS. The size distribution of exosomes was examined using a NanoSight Tracking Analysis LM20 System (NanoSight Ltd). Total RNA in exosomes were isolated using TRIzol (Invitrogen) reagent as per manufacturer's instructions. To remove DNA, the RNA was treated with RQ1 DNase (Promega, Madison, WI, USA) for 20 minutes at 37°C. The quality and quantity of the purified RNA were confirmed by measuring the absorbance at 260 and 280 nm (A 260 and A 280) using a SmartSpec Plus Spectrophotometer (Bio-Rad Laboratories, Inc., Hercules, CA, USA). RNA integrity was further verified by electrophoresis using a 1.5% agarose gel. All RNA samples were stored at -80°C for future use. RNA libraries were prepared for sequencing using standard Illumina protocols
GEO Accession
GSM4610718