Plasma/Serum Exosome and Free-Circulating RNA Isolation Kits
All-in-one system for the purification of exosomes and the sequential isolation of RNA and free-circulating RNA from plasma/serum samples
For research use only and NOT intended for in vitro diagnostics.
Version Available Here.
Plasma/Serum Exosome and Free-Circulating RNA Isolation Kits
All-in-one system for the purification of exosomes and the sequential isolation of RNA and free-circulating RNA from plasma/serum samples
Register today to receive an exclusive 15% off* on your first order.
Overview
- Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias.
- ​Versatile sample input ranges
- Isolate all sizes of free-circulating RNA, including microRNA
- The purified exosomal RNA is free from any circulating RNA-binding proteins
- No phenol extractions, Proteinase K treatment, nor carrier RNA
- No time-consuming ultracentrifugation, filtration nor special syringes are required
- No precipitation reagents, nor overnight incubation required
- Concentrate isolated exosomal RNA and are free-circulating RNA into a flexible elution volume ranging from 50 µL to 100 µL
- Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
These kits provide a fast, reliable and convenient method to sequentially isolate and concentrate exosomal RNA as well as Free-Circulating 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 as well as all sizes of the free-circulating protein-bound RNA, including microRNA. 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. Moreover, these kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. 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.
Plasma/Serum Exosome and Free-Circulating RNA Isolation Mini Kit
For sample volumes ranging from 50 µL to 1 mL.
Plasma/Serum Exosome and Free-Circulating RNA Isolation Midi Kit
For sample volumes ranging from 1 mL to 4 mL.
Plasma/Serum Exosome and Free-Circulating RNA Isolation Maxi Kit
For sample volumes ranging from 4 mL to 10 mL.
Details
Supporting Data
Figure 1. Isolation of RNA from exosomes purified from different plasma volumes. Norgen’s Plasma/Serum Exosome and Free-Circulating RNA Isolation Mini Kit (Cat# 59500) was used to isolate Exosomal RNA from different plasma volumes ranging from 0.1 mL and up to 1 mL. Two microlitres 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 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. Isolation of Free-Circulating RNA after the isolation of Exosomal RNA from different plasma volumes. Norgen’s Plasma/Serum Exosome and Free-Circulating RNA Isolation Mini Kit (Cat# 59500) was used to isolate Free-Circulating RNA after the isolation of Exosomal RNA from different plasma volumes ranging from 0.1 mL and up to 1 mL. Two microlitres of the isolated RNA was then used as the template in RT-qPCR reactions to assess the amplification of the isolated plasma free-circulating miR-21. (A) The free-circulating miR-21 is linearly decreasing with increasing the sample input volume. B) The relative amount of the free-circulating miR-21 shows excellent linearity with a percentage of recovery of more than 90%.
Figure 3. Isolation of RNA from exosomes purified from different plasma volumes. Norgen’s Plasma/Serum Exosome and Free-Circulating RNA Isolation Midi Kit (Cat# 59600) was used to isolate Exosomal RNA from different plasma volumes ranging from 1 mL and up to 4 mL. Two microlitres 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 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 4. Isolation of Free-Circulating RNA after the isolation of Exosomal RNA from different plasma volumes. Norgen’s Plasma/Serum Exosome and Free-Circulating RNA Isolation Midi Kit (Cat# 59600) was used to isolate Free-Circulating RNA after the isolation of Exosomal RNA from different plasma volumes ranging from 1 mL and up to 4 mL. Two microlitres of the isolated RNA was then used as the template in RT-qPCR reactions to assess the amplification of the isolated plasma free-circulating miR-21. (A) The free-circulating miR-21 is linearly decreasing with increasing the sample input volume. B) The relative amount of the free-circulating miR-21 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 and Free-Circulating RNA Isolation Maxi Kit (Cat# 59700) was used to isolate exosomal RNA from different plasma volumes ranging from 4 mL and up to 10 mL. Two microlitres 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 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 6. Isolation of Free-Circulating RNA after the isolation of Exosomal RNA from different plasma volumes. Norgen’s Plasma/Serum Exosome and Free-Circulating RNA Isolation Maxi Kit (Cat# 59700) was used to isolate free-circulating RNA after the isolation of exosomal RNA from different plasma volumes ranging from 4 mL and up to 10 mL. Two microlitres of the isolated RNA was then used as the template in RT-qPCR reactions to assess the amplification of the isolated plasma free-circulating miR-21. (A) The free-circulating miR-21 is linearly decreasing with increasing the sample input volume. B) The relative amount of the free-circulating miR-21 shows excellent linearity with a percentage of recovery of more than 90%.
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Kit Specifications
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| Minimum Plasma Input |
50 μL
|
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Maximum Plasma Input
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1 mL
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| 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
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| Average Yields* | Variable depending on specimen |
*Please check page 5 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. 59500 (50 preps) | Cat. 59600 (25 preps) | Cat. 59700 (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 | 2 x 20 mL | 2 x 20 mL | 2 x 20 mL |
| Lysis Additive B | 2 mL | 2 mL | 2 mL |
| Wash Solution A | 2 x 18 mL | 18 mL | 18 mL |
| Elution Solution A | 2 x 6 mL | 6 mL | 6 mL |
| Mini Filter Spin Columns | 50 | 25 | 15 |
| Mini Spin Columns | 100 | 50 | 30 |
| Collection Tubes | 100 | 50 | 30 |
| Elution Tubes (1.7 mL) | 100 | 50 | 30 |
| Product Insert | 1 | 1 | 1 |
Documentation
FAQs
Mini, Midi, Maxi
Citations
| Title | Comparative Analysis of Free-Circulating and Vesicle-Associated Plasma microRNAs of Healthy Controls and Early-Stage Lung Cancer Patients |
| Journal | Pharmaceutics. 2022. |
| Authors | Luigi Pasini, Ivan Vannini, Paola Ulivi, Michela Tebaldi, Elisabetta Petracci, Francesco Fabbri, Franco Stella and Milena Urbini |
