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.
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| Kit Specifications | |
| Minimum Plasma Input | 1 mL |
| Maximum Plasma Input | 4 mL |
| Size of RNA Purified | All sizes, including miRNA and small RNA (< 200 nt) |
| Elution Volume | 50 – 100 µL |
| Time to Complete 10 Purifications | 40 – 45 minutes |
| 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 |
These kits provide a fast, reliable and convenient spin column method for the isolation of high quality, high purity and inhibitor-free cell-free circulating DNA (cfc-DNA) from plasma/serum sample. These kits are designed to isolate all sizes of cfc-DNA from either fresh or frozen plasma/serum samples and the purified DNA is eluted into a flexible elution volume ranging from 25 µL to 50 µL. The purified plasma/serum cfc-DNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis, microarrays and NGS.
Background
Plasma/Serum cell-free circulating DNA (cfc-DNA) has the potential to provide biomarkers for certain cancers and disease states as well as fetal DNA in maternal blood. Currently, significant advancements are being made in utilizing cfc-DNA as biomarkers for the early diagnosis, prognosis and monitoring of therapy for several cancer types and autoimmune diseases. Cell-free mitocondrial DNA (cfmtDNA) is also under investigation for its clinical significance. This cfc-DNA is usually present as short fragments of less than 1000 bp. In addition, cell-free fetal DNA has been widely used as a non-invasive method for prenatal diagnosis including early identification of fetal sex, genetic studies for families at high risk for inherited genetic disorders, screening for Rhesus factor, screening for aneuploidy and identification of preeclampsia.
Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit Dx
For serum input volumes 10 µL – 200 µL. Purify high-quality DNA in 15-20 minutes
Plasma/Serum Cell-Free Circulating DNA Purification Mini Kit Dx
For serum input volumes 200 µL – 500 µL. Purify high-quality DNA in 15-20 minutes.
Plasma/Serum Cell-Free Circulating DNA Purification Midi Kit Dx
The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 25 µL to 50 µL. All components for the purification & concentration are provided in one convenient & fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here to view. For serum input volumes 1 mL – 4 mL. Purify high-quality DNA in 90 minutes.
Plasma/Serum Cell-Free Circulating DNA Purification Maxi Kit Dx
The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 25 µL to 50 µL. All components for the purification & concentration are provided in one convenient & fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here to view. For serum input volumes 5 mL – 10 mL. Two kits in one – purify and concentrate DNA from bodily fluids using a convenient two column system.
Figure 1 / 12
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| Kit Specifications | |
| Minimum Plasma/Serum Input | 10 μL |
| Maximum Plasma/Serum Input | 200 μL |
| Size of DNA Purified | ≥ 50 bp |
| Elution Volume | 25-50 μL |
| Time to Complete 10 Purifications | 15-20 minutes |
| Average Yields* | Variable depending on specimen |
*Please check page 7 in the product manual for the Plasma/Serum Average Yields and the Common DNA Quantification Methods.
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature (15-25°C) for up to 2 years without showing any reduction in performance. Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Kits contain ready-to-use Proteinase K solution, which is dissolved in a specially prepared storage buffer. The Proteinase K is stable for up to 2.5 years after delivery when stored at room temperature. To prolong the lifetime of Proteinase K, storage at 2–8°C is recommended.
| Component | Cat. Dx55100 (50 preps) | Cat. Dx55500 (50 preps) | Cat. Dx55600 (20 preps) | Cat. Dx55800 (10 preps) |
|---|---|---|---|---|
| Binding Buffer B | 2 x 40 mL | 40 mL | 2 x 85 mL 1 x 12 mL | 2 x 85 mL 1 x 40 mL |
| Proteinase K | 2 x 1 mL | 0.6 mL | 6.5 mL | 8 mL |
| Solution WN | 18 mL | 18 mL | 18 mL | 18 mL |
| Wash Solution A | 38 mL | 18 mL | 18 mL | 18 mL |
| Elution Buffer B | 8 mL | 8 mL | 30 mL | 30 mL |
| Micro Spin Columns | 50 | 50 | 20 | 10 |
| Mini Spin Columns | 50 | – | – | – |
| Midi Spin Columns | – | – | 20 | – |
| Maxi Spin Columns | – | – | – | 10 |
| Collection Tubes | 100 | 50 | 20 | 10 |
| Elution Tubes (1.7 mL) | 100 | 50 | 20 | 10 |
| Product Insert | 1 | 1 | 1 | 1 |
