Plasma/Serum RNA/DNA Purification Mini Kit
For rapid and simple sequential purification of circulating RNA, exosomal RNA and cell-free circulating DNA from plasma/serum samples
For research use only and NOT intended for in vitro diagnostics.
Plasma/Serum RNA/DNA Purification Mini Kit
For rapid and simple sequential purification of circulating RNA, exosomal RNA and cell-free circulating DNA from plasma/serum samples
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Features and Benefits
- Isolate all sizes of circulating and exosomal RNA, including microRNA
- Isolate all sizes of circulating DNA from plasma and serum samples
- Isolate viral and bacterial DNA and RNA
- Versatile plasma and serum input volumes (10 µL - 200 µL)
- No phenol extractions
- Bind and elute all RNA irrespective of size or GC content, without bias
- Concentrate circulating RNA, exosomal RNA and cell-free circulating DNA into a flexible elution volume ranging from 10 µL to 25 µL
- Isolate inhibitor-free nucleic acids
- Purify high-quality RNA and DNA in 30 minutes
- Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen's NGS services
- Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
Norgen's Plasma/Serum RNA/DNA Purification Mini Kit provides a fast, reliable, reproducible and simple procedure for the sequential isolation of circulating RNA, exosomal RNA and Cell-Free Circulating DNA (cfc) from the same Plasma/Serum sample. It can sequentially isolated RNA and DNA from small plasma/serum input ranging from 10 µL to 200 µL. The kit is designed to isolate all sizes of circulating RNA, including microRNA, all sizes of exosomal RNA as well as all sizes of cfc-DNA from fresh or frozen plasma or serum samples. Norgen's Plasma/Serum RNA Purification Kit provides a clear advantage over other available kits in that it does not require Phenol/Chloroform or any Protease treatments for the isolation of plasma/serum RNA or DNA . RNA and DNA can be eluted into a flexible elution volume ranging from 10 µL to 25 µL. Purified RNA can be used in a number of sensitive downstream applications including reverse transcription qPCR, reverse transcription PCR, NGS, Northern blotting, RNase protection and primer extension, and expression array assays. Purified DNA can be used in a number of sensitive downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis.
Background
Typical yields of free-circulating, exosomal RNA and cfc-DNA vary depending on the input sample, as the amount of RNA and/or DNA present in plasma and serum will depend upon the health status of the individual. Normally, the RNA/DNA yield from plasma or serum is highly variable (range from 1 to 100 ng/mL). Variability is also observed between samples collected from the same donor at different times during the day. This kit is suitable for the isolation of RNA/DNA from 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.
Details
Supporting Data
Figure 1. Purification of Circulating RNA from Different Plasma Volumes. Norgen's Plasma/Serum RNA/DNA Purification Mini Kit was used to purify circulating RNA from 50 µL, 100 µL and 200 µL plasma prepared from blood collected on EDTA. Three microlitres of the purified RNA was then used as the template in RT-qPCR reactions to detect miR-21 (Figure 1A) and the housekeeping 5S rRNA transcript (Figure 1B). The relative amount of both the miR-21 (Figure 1A) and the 5S rRNA transcript (Figure 1B) is linearly increasing with increasing the sample input volume.
Figure 2. Purification of Cell-Free Circulating DNA from Different Plasma Volumes. Norgen's Plasma/Serum RNA/DNA Purification Mini Kit was used to purify cell-free circulating DNA from 50 µL, 100 µL and 200 µL plasma prepared from blood collected on EDTA. Three microlitres of the purified DNA was then used as the template in qPCR reactions to detect the housekeeping 5S rRNA transcript. The average Ct value for the 5S rRNA gene is linearly decreasing with increasing the sample input volume.
Figure 3. Eluting Purified Circulating RNA from into Different Elution Volumes. Norgen's Plasma/Serum RNA/DNA Purification Mini Kit was used to purify circulating RNA from 200 µL plasma prepared from blood collected on EDTA and eluted in 10 µL, 15 µL, and 25 µL. Three microlitres of the purified RNA was then used as the template in RT-qPCR reactions to detect miR-21 (Figure 3A) and the housekeeping 5S rRNA transcript (Figure 3B). The relative amount of both the miR-21 (Figure 3A) and the 5S rRNA transcript (Figure 3B) is decreasing with increasing the elution volume indicating the efficient concentration of the plasma circulating RNA in a very low elution volume.
Figure 4. Eluting Purified Cell-Free Circulating DNA from into Different Elution Volumes. Norgen's Plasma/Serum RNA/DNA Purification Mini Kit was used to purify circulating cell-free circulating DNA from 200 µL plasma prepared from blood collected on EDTA and eluted in 15 µL, 25 µL and 50 µL. Three microlitres of the purified cell-free circulating DNA was then used as the template in qPCR reactions to detect the housekeeping 5S rRNA gene. The relative amount of the 5S rRNA gene is decreasing with increasing the elution volume indicating the efficient concentration of the plasma circulating cell-free circulating DNA in a very low elution volume.
Figure 5. Effective and Consistent Detection of miRNA from Plasma. Norgen's Plasma/Serum RNA/DNA Purification Mini Kit can effectively isolate miRNA from plasma. Circulating miRNA was isolated from 200 µL plasma using Norgen's Plasma/Serum RNA/DNA Purification Mini Kit, competitor Q's kit and competitor E's kit. Circulating miRNA was isolated from 600 µL using competitor A's kit. Stem loop RT-qPCR using primers specific to miR-21 was performed. In brief, 1 microliter of the 15 µL purified RNA using Norgen's Plasma/Serum RNA/DNA Purification Mini Kit, competitor Q's kit and 3.3 microliters of the 50 µL purified RNA using competitor E's kit and competitor A's kit was then subjected to a 20 µL reverse transcription using miR-21 stem-loop reverse primer. Three microliters of the reverse transcription was used in a 20 µL real-time PCR reaction with primers to detect the human miR-21. Norgen's Plasma/Serum RNA Purification Kit is the only product that showed the most consistent and the highest recovery of the miR-21 transcripts as compared to the other isolation methods. The recovery of the miRNA from 200µL plasma was higher than that recovered from RNA purified from 600µL using competitor A's kit.
Figure 6. Effective and Consistent Detection of Cell-Free Circulating DNA from Plasma. Norgen's Plasma/Serum RNA/DNA Purification Mini Kit can effectively isolate cell-free circulating DNA from plasma. Cell-Free Circulating DNA was isolated from 200 µL plasma using Norgen's Plasma/Serum RNA/DNA Purification Mini Kit, competitor Q's kit and competitor M's kit. Three microliters of the purified DNA was used in a 20 µL real-time PCR reaction with primers to detect the housekeeping 5S rRNA gene. Norgen's Plasma/Serum Cell-Free Circulating DNA Purification Kit is the only product that showed the most consistent and the highest recovery of the housekeeping 5S rRNA gene as compared to the other isolation methods.
Figure 7. Determination of the Amount of Inhibition Present in Plasma Cell-Free Circulating DNA Samples when Detecting the Human 5S Gene. DNA was isolated from 0.2 mL of plasma using Norgen's Plasma/Serum RNA/DNA Purification Mini Kit, competitor Q's kit and competitor M's kit. Increasing volumes of the elution (3, 6 and 9 μL) were used in a 20 μL qPCR reaction to observe any increase in Ct value. An increase in Ct value with increasing amount of template would be a clear indication of PCR inhibitors present in the sample. An increase in elution volume used in the PCR did not greatly affect the Ct value generated from Norgen samples, however inhibition was observed when 9 μL of competitor Q's kit elution was used as the template.
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Kit Specifications
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Minimum Plasma/Serum Input Volume
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10 μL
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Maximum Plasma/Serum Input Volume
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200 μL
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| Size of RNA Purified |
All sizes including small RNA (<200 nt)
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| Size of DNA Purified |
≥ 50 bp
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| Minimum Elution Volume |
10 μL
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| Maximum Elution Volume | 25 μL |
| Time to Complete 10 Purifications |
15-20 minutes
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| Average Yield |
Variable depending on specimen
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Note: Do not exceed the recommended sample input volume of 200 µL.
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.
Component | Cat. 55200 (50 preps) |
|---|---|
Lysis Buffer A | 30 mL |
Wash Solution A | 38 mL |
Solution WN | 18 mL |
Elution Solution A | 6 mL |
Elution Buffer B | 8 mL |
Micro Spin Columns | 50 |
Micro-Elute RNA Spin Columns | 50 |
Collection Tube | 100 |
Elution Tubes (1.7 mL) | 100 |
Product Insert | 1 |
Documentation
FAQs
Mini
Citations
| Title | A straightforward method to quantify circulating mRNAs as biomarkers of colorectal cancer |
| Citation | Scientific Reports 2023. |
| Authors | Marie Grosgeorges, Laurence Picque Lasorsa, Brice Pastor, Corinne Prévostel, Evelyne Crapez, Cynthia Sanchez, Florence Frayssinoux, Marta Jarlier, Véronique Pezzella, Laure Monard, Marc Ychou, Alain R. Thierry, Thibault Mazard & Philippe Blache |
