Quick and easy protocol for the isolation of exosomal RNA from exosomes previously purified using Norgen's Exosome Purification Kits, ultracentrifugation, size-exclusion chromatography or most other commercial methods. This method also enables the sequential isolation of free-circulating RNA from various sample types.
Details
Supporting Data
Figure 1. Isolation of RNA from exosomes purified from different urine volumes.
Norgen's Exosomal RNA Isolation Kit (Cat# 58000) was used to isolate RNA from exosomes isolated from different urine volumes purified using Norgen's Urine Exosome Purification Kits (Cat# 57700, 57800 and 57900). 2 μL of the isolated RNA was then used as the template in RT-qPCR reactions to assess the amplification of the isolated urinary exosomal miR-30a. (A) The average Ct value for urinary exosomal miR-30a is linearly decreasing with increasing the sample input volume. B) The relative amount of the urinary exosomal miR-30a shows excellent linearity with a percentage of recovery of more than 90%.
Figure 2. Isolation of RNA from exosomes purified from different plasma volumes.
Norgen's Exosomal RNA Isolation Kit (Cat# 58000) was used to isolate RNA from exosomes isolated from different plasma volumes purified using Norgen's Plasma/Serum Exosome Purification Kits (Cat# 57400, 57500 and 57600). 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 the 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%.
Kit Specifications | |
Sample type | Purified exosomes |
Size of RNA Purified | All sizes, including miRNA and small RNA (< 200 nt) |
Elution Volume | 50-100 μL |
Time to Complete 10 Purifications | 35-45 minutes |
Average Yield | Depends on specimen |
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
Component | Cat. 58000 (50 preps) |
|---|---|
Lysis Buffer A | 20 mL |
Lysis Additive B | 2 mL |
Wash Solution A | 38 mL |
Elution Solution A | 6 mL |
Mini Spin Columns | 50 |
Collection Tubes | 50 |
Elution Tubes (1.7 mL) | 2 x 25 |
Product Insert | 1 |
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