Plasma/Serum Exosome Purification and RNA Isolation Kits
All-in-one system for the purification of exosomes and the subsequent isolation of RNA from plasma/serum samples
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For research use only and NOT intended for in vitro diagnostics.
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Plasma/Serum Exosome Purification and RNA Isolation Kits
All-in-one system for the purification of exosomes and the subsequent isolation of RNA from plasma/serum samples
Overview
- Purification and enrichment of intact plasma/serum exosomes for functional studies
- Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias
- Versatile plasma/serum input volume
- No phenol extractions, Proteinase K treatment, nor carrier RNA required
- No time-consuming ultracentrifugation, filtration nor special syringes required
- No precipitation reagents, nor overnight incubation required
- Compatible with plasma/serum from most species
- Pure exosomes are purified and are free from any other RNA-binding proteins
- Purification is based on spin column chromatography that uses Norgen’s proprietary resin
- The purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen's NGS services
Norgen’s Plasma/Serum Exosome and RNA Isolation Kits constitute all-in-one systems for the purification of exosomes and the sequential isolation of exosomal RNA from different plasma/serum sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias. These kits provide a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. The purified RNA can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.
Norgen’s Plasma/Serum Exosome and RNA Isolation Mini Kit
For sample volumes ranging from 50 µL to 1 mL. This kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL.
Norgen’s Plasma/Serum Exosome and RNA Isolation Midi Kit
For sample volumes ranging from 1 mL to 4 mL. This kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL.
Norgen’s Plasma/Serum Exosome and RNA Isolation Maxi Kit
For sample volumes ranging from 4 mL to 10 mL. This kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL.
Details
Supporting Data
Figure 1. Isolation of RNA from exosomes purified from different plasma volumes. Norgen's Plasma/Serum Exosome Purification and RNA Isolation Mini Kit (Cat# 58300) 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 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%.
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%.
Kit Specifications
|
|
Minimum Plasma/Serum Input |
50 μL
|
Maximum Plasma/Serum Input
|
1 mL
|
Size of Exosomes Purified
|
40 nm - 150 nm
|
Size of RNA Purified | All sizes, including miRNA and small RNA (< 200 nt) |
Elution Volume | 50-100 μL |
Time to Complete 10 Purifications |
35 - 40 minutes
|
Average Yields* | Variable depending on specimen |
*Please check page 4 of the product insert for the average yields and the common RNA quantification methods.
Storage Conditions and Product Stability
All solutions 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.
Important Note
This kit is suitable for the purification of exosomes from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.
Component | Cat. 58300 (50 preps) | Cat. 58500 (25 preps) | Cat. 58600 (15 preps) |
---|---|---|---|
Slurry E | 12.5 mL | 12.5 mL | 12.5 mL |
ExoC Buffer | 8 mL | 8 mL | 2 x 8 mL |
ExoR Buffer | 12 mL | 12 mL | 12 mL |
Lysis Buffer A | 20 mL | 20 mL | 20 mL |
Lysis Additive B | 2 mL | 2 mL | 2 mL |
Wash Solution A | 38 mL | 18 mL | 18 mL |
Elution Solution A | 6 mL | 6 mL | 6 mL |
Mini Filter Spin Columns | 50 | 25 | 15 |
Mini Spin Columns | 50 | 25 | 15 |
Collection Tubes | 50 | 25 | 15 |
Elution Tubes (1.7 mL) | 100 | 50 | 30 |
Product Insert | 1 | 1 | 1 |
Documentation
FAQs
Mini, Midi, Maxi
Citations
Mini (58300)
Title | Cancer cells exploit an orphan RNA to drive metastatic progression |
Journal | Nature Medicine. 2018. |
Authors | Lisa Fish, Steven Zhang, Johnny X. Yu, Bruce Culbertson, Alicia Y. Zhou, Andrei Goga & Hani Goodarzi |
Maxi (58500)
Title | Small RNA-sequence analysis of plasma-derived extracellular vesicle miRNAs in smokers and patients with chronic obstructive pulmonary disease as circulating biomarkers |
Journal | Journal of Extracellular Vesicles, 8(1). 2019. |
Authors | Sundar, I. K., Li, D., & Rahman, I. |