These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples 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. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS.
Cell-free circulating RNA, including exosomal RNA in plasma or serum, has the potential to provide biomarkers for certain cancers and disease states, and includes tumor-specific extracellular RNA in the blood. Exosomes are 40 – 100 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. The exosomes contain cell-specific proteins, lipids and RNAs, which are transported to other cells, where they can alter function and/or physiology. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. Recent work has demonstrated the presence of distinct subsets of microRNAs within exosomes which depend upon the tumour cell type from which they are secreted. For this reason, exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses, they can be efficiently recovered from biological fluids, such as plasma or serum.
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 μL to 200 μL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 μL to 25 μL.
This utilizes a two-column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 μL to 1.5 mL. 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 50 μL to 100 μL.
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. 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 50 μL to 100 μL.
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| Kit Specifications | |
| Sample Type | Plasma/Serum |
| Anti-Coagulant (for Plasma)† | EDTA or Citrate |
| Sample Volume Range | 250 μL to 1.5mL |
| Minimum Elution Volume | 50 μL |
| Maximum Elution Volume | 100 μL |
| Time to Complete 10 Purifications | 35 – 40 minutes |
| Size of RNA Purified | All sizes, including miRNA and small RNA (<200 nt) |
| Average Yields¥ | Variable depending on specimen |
† This kit is suitable for the isolation of RNA 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
¥ Please check page 6 for Average Plasma/Serum Yields and 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.
| Component | Cat. 73700 (50 preps) | Cat. 73710 (20 preps) | Cat. 73720 (10 preps) |
|---|---|---|---|
| Lysis Buffer A | 2 x 20 mL | 100 mL | 1 x 130 mL 1 x 30 mL |
| Wash Solution A | 18 mL* | 1 x 38 mL* 1 x 18 mL* | 38 mL* |
| Elution Solution A | 6 mL | 6 mL | 6 mL |
| Elution Buffer F | – | – | 15 mL |
| EXTRACLean Micro Spin Columns | 50 | – | – |
| EXTRACLean Mini Spin Columns | – | 20 | 10 |
| EXTRACLean Midi Spin Columns | – | 20 | – |
| EXTRACLean Maxi Spin Columns | – | – | 10 |
| Collection Tubes | – | 20 | 10 |
| Elution Tubes (1.7 mL) | 50 | 20 | 10 |
| Product Insert | 1 | 1 | 1 |
Norgen’s Saliva DNA Collection and Preservation Devices are designed for 1) simple and non-invasive saliva collection and 2) preservation of DNA in saliva samples at ambient temperature. Each of the 50 Saliva DNA Collection and Preservation Devices consists of 3 components: (1) Saliva Collection Funnel and Collection Tube, (2) Collection Tube Cap, and (3) Norgen’s Saliva DNA Preservative contained within a sealed squeezable ampoule. Saliva samples are collected by spitting inside the Collection Funnel which has been assembled with the Collection Tube. After collecting the required volume of saliva the Collection Funnel is removed and the contents of the Preservative Ampoule are then added and mixed with the collected saliva. The Saliva Collection Tube is subsequently sent to the laboratory for DNA isolation and analysis. DNA can be isolated from the preserved saliva samples using Norgen’s Saliva DNA Isolation Kit (Cat# Dx45400). Each of Norgen’s Collection Tubes is labeled with a unique serial number that can be used for secure and anonymous tracking of the sample. The saliva DNA in preserved samples is stable for more than 2 years at room temperature. This kit is ideal for collecting and preserving DNA samples for in vitro diagnostic use for medical purposes.
Norgen’s Saliva DNA Preservative is an aqueous storage buffer designed for rapid cellular lysis and subsequent preservation of DNA from fresh specimens. The buffer prevents the growth of Gram-negative and Gram-positive bacteria and fungi, and also inactivates viruses allowing the resulting non-infectious samples to be handled and shipped safely. In addition, the buffer eliminates the need to immediately process or freeze samples and allows the samples to be shipped to centralized testing facilities at ambient temperature. The components of the buffer allow samples to be stored for more than 2 years without any detectable DNA degradation.
Prior to saliva DNA isolation, vortex the Collection Tube containing preserved saliva for 10 seconds and incubate at 55°C for one hour. Saliva DNA can now be isolated from the preserved saliva samples using Norgen’s spin-column based Saliva DNA Isolation Kit Dx (Cat# Dx45400) or other commercially available methods that have been validated for use with Norgen’s Saliva DNA Collection and Preservation Devices.
NOTE: This product is not available for sale in the United States.
Figure 1 / 3
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| Kit Specifications | |
| Volume of Saliva Collected | 2 mL |
| Volume of Saliva-Preservative Mix | 4 mL |
| Preservation Temperature | Room Temperature |
| Preservation time | Over 2 years at room temperature |
Shelf Life and Handling
| Kit Components | Cat. 49000 (50 Devices) |
|---|---|
| Individual Saliva DNA Collection and Preservation Devices | 50 |
| Donor Procedure Flowchart | 1 |
| Product Insert | 1 |
| Individual Saliva DNA Collection and Preservation Device Contents | |
| Saliva Collection Funnel and Collection Tube | 1 |
| Collection Tube Cap | 1 |
| Preservative Ampoule | 1 |
| Donor Instructions | 1 |
Norgen’s RNA purification kits isolate total RNA with minimal amounts of genomic DNA contamination. However, for some sensitive downstream applications, it may be desirable to remove all traces of residual DNA. Norgen’s RNase-free DNAse I Kit, with Enzyme Incubation Buffer, can be used for optional on-column DNase digestion with any of Norgen’s RNA purification kits. Alternatively, after isolating total RNA using one of Norgen’s RNA purification kits, the RNA elution can be treated with this DNase I. The RNA can then be purified from the DNase using Norgen’s RNA Clean-Up and Concentration Kit (Cat# 23600), and the RNA can then be used in downstream applications.
Each RNase-Free DNase I Kit is supplied complete with sufficient enzyme and enzyme incubation buffer for 50 or 200 reactions.
Storage Conditions
The DNase I provided is in lyophilized form. It is stable for at least 3 months if stored at room temperature. However, it is recommended to store the DNase I vial at 2 – 8ºC (or below) upon receipt to maintain stability beyond 3 months. Buffer DR and Enzyme Incubation Buffer can be stored at room temperature. After reconstitution with Buffer DR (see product manual), the DNase I should be stored at -20ºC. All reagents should remain stable for at least 1 year in their unopened containers at the appropriate storage temperature.
| Component | Cat. 25710 (50 rxns) | Cat. 25720 (200 rxns) |
|---|---|---|
| DNase I | 1 vial / 1,600 units | 4 vials (1,600 units/vial) |
| Buffer DR | 1 mL | 4 x 1 mL |
| Enzyme Incubation Buffer | 6 mL | 4 x 6 mL |
| Product Insert | 1 | 1 |
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