Figure 2. Comparing exosome yields from different purification methods. Intact exosomes were purified from 1 mL plasma using Norgen's Plasma/Serum Exosome Purification and RNA Isolation Mini Kit (Cat# 58300), Competitor S's kit and Competitor L's kit (from plasma). Exosomes purified using Norgen's kit were resuspended in 200 µL of Norgen's ExoR buffer, diluted 1:1,000 and visualized on the NanoSight LM10 instrument. The analysis shows that Norgen's kit isolated 55 nm exosomes with a recovery of 9.64 x 10¹³ particles/mL plasma. No impurities were found to be contaminating the exosomes purified using Norgen's Plasma/Serum Exosome Purification and RNA Isolation Kit. Additionally, exosomes with a broader size range covering from 50 nm - 150 nm were purified from 1 mL plasma with a higher concentration compared to the other two methods.

Figure 3. NanoSight data showing intact exosomes purified from 1 mL and 10 mL plasma. Intact exosomes were purified from 1 mL plasma using Norgen's Plasma/Serum Exosome Purification and RNA Isolation Mini Kit (Cat# 58300) and from 10 mL plasma using Norgen's Plasma/Serum Exosome Purification and RNA Isolation Maxi Kit (Cat# 58600). Exosomes purified using Norgen's mini kit were resuspended in 200 µL of Norgen's ExoR buffer whereas exosomes purified using Norgen's Maxi kit were resuspended in 600 µL Norgen's ExoR buffer, diluted 1:1,000 and visualized on the NanoSight LM10 instrument. The analysis shows that the purification of exosomes is linear as 4.04 x 10¹⁰ particles/mL were recovered from 1 mL plasma, whereas 2.95 x 10¹¹ particles/mL were recovered from 10 mL plasma.

Figure 4. Isolation of RNA from exosomes purified from different plasma volumes. Norgen’s Plasma/Serum Exosome Purification and RNA Isolation Midi Kit (Cat# 58500) was used to isolate RNA from exosomes purified from different plasma volumes using the same kit. 2 μL of the isolated RNA was then used as the template in RT-qPCR reactions to assess the amplification of the isolated plasma exosomal miR-26a. (A) The average Ct value for plasma exosomal miR-26a is linearly decreasing with increasing sample input volume. B) The relative amount of the plasma exosomal miR-26a shows excellent linearity with a percentage of recovery of more than 90%.

Figure 5. Isolation of RNA from exosomes purified from different plasma volumes. Norgen’s Plasma/Serum Exosome Purification and RNA Isolation Maxi Kit (Cat# 58600) was used to isolate RNA from exosomes purified from different plasma volumes using the same kit. 2 μL of the isolated RNA was then used as the template in RT-qPCR reactions to assess the amplification of the isolated plasma exosomal miR-26a. (A) The average Ct value for plasma exosomal miR-26a is linearly decreasing with increasing sample input volume. B) The relative amount of the plasma exosomal miR-26a shows excellent linearity with a percentage of recovery of more than 90%.