Centrifuge 5810 no 0012529 rotor a 4 81

Blood samples consisted of 19 mL of whole blood collected into ethylenediaminetetraacetic acid tubes within 48 h of enrollment in the SMC-ROCI. Samples were centrifuged at 480× g (Eppendorf Centrifuge 5810 No. 0012529-rotor A-4-81) for 10 min at 4 °C within 4 h of collection. Several plasma aliquots from each study participant were then isolated and stored at −80 °C for further analysis.

This was a Mayo Clinic Institutional Review Board (Mayo Clinic IRB# 15-006351) approved and Health Insurance Portability and Accountability Act (HIPAA) compliant study. Written informed consent was obtained from all patients. Nineteen samples were obtained from glioma patients undergoing surgery (6 females ages 35–67; 13 male ages 27–73). Nineteen anonymized control samples from normal donors were obtained through discarded material from the Mayo Clinic (Rochester) Blood Bank. All samples were acquired through collection of whole blood in ethylenediaminetetraacetic acid (EDTA) tubes. After blood collection, the samples were spun at 3,000 RPM or 1,811 xg (Eppendorf Centrifuge 5810 No. 0012529- rotor A-4-81) for 10 min. Plasma isolated from blood was then transferred into a 15 ml conical tube (Falcon No. 352097) and spun at 3,000 RPM (1,811 xg) for 15 min (Figure 1A). Plasma (1–9 ml) was recovered from each sample and stored in a sterile cryogenic vial (Corning Incorporated No. 430488) at −20°C.

Blood samples (≥ 15 mL) were collected for plasma preparation. Plasma was prepared by drawing peripheral blood into ethylenediaminetetraacetic acid tubes, followed by centrifugation at 480 g (Eppendorf Centrifuge 5810 No. 0012529-rotor A-4-81) for 10 min at 4 °C. Several plasma aliquots from each study participant were isolated and stored at − 80 °C until further analysis. Prior to exosome isolation and RNA purification, plasma samples were pre-filtered using 0.8 µm syringe filters, followed by additional centrifugation to eliminate residual cellular debris.