Ultracentifugation and nanoscale deterministic lateral displacement (nanoDLD) of samples for exRNA analysis

An integrative analysis of human biofluid data in the exRNA Atlas revealed the existence of distinct extracellular RNA cargo types. To gain further insight on the biological nature of these cargo types, we correlated exRNA Atlas cargo profiles with a variety of other RNA-seq profiles. This study focuses on those samples obtained via ultracentrifugation and nanoscale deterministic lateral displacement (nanoDLD). Whole blood samples (2 to 5 ml) were collected by the team of Dr. Ashutosh Tewari at the Icahn School of Medicine at Mount Sinai, New York, Department of Urology by venipuncture from 9 consenting adult male Prostate Cancer patients under Institute Review Board approved protocols (GCO # 06-0996, 14-0318, and surgical consent) in purple capped tubes.  After blood collection, serum was isolated using BD Vacutainer blood collection tubes, serum separation tubes (Fisher Scientific, Cat # 368016) and kept at -80°C until further steps were taken for exosome isolation. Serum was rapidly thawed prior to EV isolation with both nanoDLD and UC.