cf-DNA/cf-RNA Preservative Tubes Dx
For Whole Blood Collection and Preservation of Cell-Free Circulating DNA and RNA
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Intended for in vitro diagnostics use.
Research version available here
This product is not available for purchase in Canada, or the United States.
cf-DNA/cf-RNA Preservative Tubes Dx
For Whole Blood Collection and Preservation of Cell-Free Circulating DNA and RNA
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Features and Benefits
- CE-IVD marked in accordance with EU Directive 98/79/EC
- Ideal for use in in vitro diagnostic workflows
- Fixative-free preservative, no cross-linking of DNA
- Preserve cf-DNA/ct-DNA for 30 days at ambient temperature and for up to 8 days at 37°C.
- Preserve cf-RNA for 30 days at ambient temperature
- Preserve Circulating Tumour Cells (CTCs) for 14 days at ambient temperature
- No plasma volume loss after shipping/transportation
- Prevent hemolysis allowing better separation of plasma
- Prevent apoptosis of blood cells and fragmentation of genomic DNA
- Produce high quality/quantity of plasma cf-DNA/ct-DNA/cf-RNA
- Vacuumed to draw 8.4 mL of blood in 10 mL tubes
- The procedure for plasma isolation using this kit is compliant with ISO 20186-3:2019
Norgen's cf-DNA/cf-RNA Preservative Tubes Dx are closed, evacuated plastic tubes for the collection and the preservation of cf-DNA, circulating tumor DNA, cf-RNA and circulating tumor cells in human whole blood samples during storage and shipping. Using these tubes in combination with Norgen's Plasma/Serum Cell-Free Circulating DNA Purification Kits or Norgen's Plasma/Serum RNA Purification Kits enables the purification and concentration of an inhibitor-free cf-DNA/ct-DNA/cf-RNA in a very small elution volume.
These tubes are ideal for collecting and preserving cf-DNA and cf-RNA samples for in vitro diagnostic use for medical purposes.
Performance
Norgen's cf-DNA/cf-RNA Preservative Tubes Dx abridge the collection/processing of whole blood for the subsequent purification of cf-DNA and/or cf-RNA. The cf-DNA/cf-RNA Preservative Tubes Dx preserve and inhibit the programmed cell-death (apoptosis) of the blood, hence preventing the release of intracellular DNA/RNA into plasma. Whole blood samples can be shipped and stored at room temperature for the subsequent processing of plasma. The cf/ct DNA and cf-RNA levels are stable for up to 30 days at room temperature. The cf-DNA is also stable for up to 8 days at 37°C.
Workflow
NOTE: This product is not available for sale in the United States. For the research use only version, see Cat. 63950
Details
Supporting Data
Figure 1. Effect of ambient temperature storage on cf-DNA (pDNA), exemplified by the short Alu (115bp) fragment, and genomic DNA (gDNA), exemplified by the large Alu (247 bp) fragment. Blood samples were drawn into either: 1) EDTA tubes, 2) Competitor tubes or 3) Norgen's cf-DNA/cf-RNA Preservative Tubes Dx and stored at room temperature. Aliquots of blood were removed at the indicated times and the plasma was separated. DNA was isolated, and pDNA and gDNA concentrations were determined by real-time PCR using a short ALU gene target (115 bp) representing the pDNA and a long ALU gene (247 bp) representing the gDNA. Levels of these two fragments should stay the same for the duration indicating stabilization and no hemolysis. As expected, there was no stabilization of cf-DNA and extensive hemolysis in the EDTA tube. Competitor showed significantly lower Ct values for both genes after 14 days, whereas cf-DNA was stable for 30 days at room temperature for Norgen's cf-DNA/cf-RNA Preservative Tubes Dx.
Figure 2. No plasma volume loss after shipping/transportation. Blood was drawn from 6 different donors in duplicate. One set was kept in the lab at room temperature and the other was packed in an insulated box and shipped from Thorold, ON via overnight air freight to Winnipeg, MB and then back to Thorold ON (elapsed time 72 h). Upon return, preserved samples were stored at room temperature for 7 days before plasma was separated. The plasma volume recovered from Norgen's cf-DNA/cf-RNA Preservative Tubes Dx did not change before shipping or after shipping (6-7 mL recovered plasma). For both Competitor tubes and EDTA tubes the plasma volume recovered before shipping was ~ 4 mL and after shipping was ~ 2.5 mL.
Figure 3. Hemolysis of collected blood measured over time. Blood samples were drawn into: 1) EDTA tubes, 2) Competitor tubes and 3) Norgen's cf-DNA/cf-RNA Preservative Tubes Dx and stored for up to 30 days. Hemolysis was determined by measuring the absorption of free hemoglobin in plasma from 3 subjects at 414 nm over several time points. Mean absorption is shown. The amount of free hemoglobin increased rapidly with each additional storage day in the EDTA tubes and Competitor tubes, and remained relatively constant in Norgen's cf-DNA/cf-RNA Preservative Tubes Dx.
Figure 4. Prevent cell lysis and the release of gDNA and accumulation of apoptotic ladder in plasma. Blood samples drawn into three different tubes (Norgen's cf-DNA/cf-RNA Preservative Tubes Dx, EDTA, and Competitor) and stored for up to 30 days. Norgen's cf-DNA/cf-RNA Preservative Tubes Dx helps prevent the release of high molecular weight gDNA into plasma while also minimizing the accumulation of contaminating apoptotic ladder from dying peripheral blood leukocytes. As indicated in the black circle, gDNA contamination is at a very low level as compared to both the Competitor and EDTA tubes. This is indicative of very low levels of cell lysis and subsequent release of gDNA into the preserved plasma sample.
Figure 5. Effect of high temperature (37°C) storage for 8 days. Blood samples were drawn into either EDTA tubes, Competitor tubes or Norgen's cf-DNA/cf-RNA Preservative Tubes Dx and stored at 37°C. cf-DNA was then isolated from processed plasma. gDNA concentration was determined by real-time PCR using a long ALU gene target (247 bp). The gDNA - target in EDTA tubes showed a significant drop in the Ct. value after 1 day of storage at 37°C compared to the initial time point, and continued to drop until the 8th day of storage. For Competitor, the gDNA - target remained stable up to the 4th day of storage and then the Ct. values started to significantly drop, indicating poor stabilization beyond day 4. Norgen's cf-DNA/cf-RNA Preservative Tubes stabilized samples for 8 days.
Figure 6. High Quantity of cf-DNA from Plasma preserved using Norgen's cf-DNA/cf-RNA Preservative Tubes Dx. Blood samples from the same donor were drawn into either Competitor tubes or Norgen's cf-DNA/cf-RNA Preservative Tubes Dx and stored at room temperature for 7 days. Norgen's cf-DNA/cf-RNA preservative Tube Dx recovered 6.5mL plasma whereas the Competitor's tube recovered 3.5mL plasma. The cf-DNA was then isolated from the entire plasma volume recovered from each tube using Norgen's Plasma/Serum Cell-Free Circulating DNA Purification Midi Kit (Cat. 55600) and Norgen's Plasma/Serum Cell-Free Circulating DNA Purification Maxi Kit (Cat. 55800). The quantity of DNA recovered from both samples were assessed using the Agilent Bio-analyzer High Sensitivity DNA Chip. As can be seen in the bioanalyzer trace, Norgen's preservative tube yielded more cf-DNA (Blue peak) as compared to the cf-DNA recovered from the Competitor's preservative tube.
Figure 7. Effect of ambient temperature storage on cf-RNA, exemplified by the 18S rRNA transcript, HPRT1 mRNA transcript and miR-21. Blood samples were drawn into either: 1) Competitor tubes or 2) Norgen's cf-DNA/cf-RNA Preservative Tubes Dx. Competitor’s tubes were stored at room temperature for 14 days whereas Norgen’s cf-DNA/cf-RNA Preservative Tubes Dx were stored for 30 days at room temperature. At the indicated times each tube was processed and the plasma was separated. Cf-RNA was isolated and the stability of the purified cf-RNA was determined by rt-qPCR amplification targeting the 18S rRNA transcript, HPRT1 mRNA transcript and miR-21. Levels of these three targets should stay the same for the duration indicating stabilization. Competitor showed significantly higher Ct values for the three targets after 7 days indicating cf-RNA degradation, whereas cf-RNA was stable for 30 days at room temperature for Norgen's cf-DNA/cf-RNA Preservative Tubes Dx.
Kit Specifications
|
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Tube Size
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16 x 100 mm
|
Tube Type | PET Tube |
Blood Draw Volume
|
8.4 mL in 10 mL Tubes
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Preservative Volume |
1.6 mL
|
Recovered Plasma Volume | Approximately 6-7 mL (not affected by shipping) |
Anticoagulant |
Proprietary
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Preservative |
Non formaldehyde - Proprietary preservation agent
|
Length of cf-DNA Preservation |
Up to 30 days at room temperature (15-25°C)
Up to 8 days at 37°C |
Length of cf-RNA Preservation |
Up to 30 days at room temperature (15-25°C)
|
Length of CTCs Preservation |
Up to 14 days at room temperature (15-25°C)
|
Storage Conditions and Product Stability
- When stored at 15-30°C, unfilled Norgen cf-DNA/cf-RNA Preservative Tubes Dx are stable through the expiration date. See tube label for expiry date.
- Short-term storage/shipping at -20°C to 37°C for up to 10 days is acceptable for unfilled Norgen cf-DNA/cf-RNA Preservative Tubes Dx.
- Do not freeze unfilled Norgen cf-DNA/cf-RNA Preservative Tubes Dx for long term storage over 10 days. Freezing unfilled tubes may lead to loss of vacuum and precipitation may occur.
- Cell-free DNA in blood samples collected in Norgen's cf-DNA/cf-RNA Preservative Tubes Dx is stable for up to 30 days when stored at room temperature (15-25°C)
- Cell-free DNA in blood samples collected in Norgen's cf-DNA/cf-RNA Preservative Tubes Dx is stable for up to 8 days when stored at 37ºC.
- Cell-free RNA in blood samples collected in Norgen's cf-DNA/cf-RNA Preservative Tubes Dx is stable for up to 30 days when stored at room temperature (15-25°C)
- Circulating tumor cells (CTCs) in blood samples collected in Norgen's cf-DNA/cf-RNA Preservative Tubes Dx are stable for up to 14 days at room temperature (15-25ºC).
Component | Cat. Dx63950 (50 tubes) |
---|---|
cf-DNA/cf-RNA Preservative Tubes Dx | 50 |
Product Insert | 1 |
Flowchart | 1 |
Documentation
Citations
Title | Response to BRAF/MEK Inhibition in A598_T599insV BRAF Mutated Melanoma |
Journal | Case Reports in Oncology.2019. |
Authors | Sara Bjursten, Christoffer Vannas, Stefan Filges, Florian Puls, Ankur Pandita, Henrik Fagman, Anders Ståhlberg, and Max Levin |
Title | Evaluation of Storage Tubes for Combined Analysis of Circulating Nucleic Acids in Liquid Biopsies |
Journal | International Journal of Molecular Sciences. 2019. |
Authors | Aoife Ward Gahlawat, Judith Lenhardt, Tania Witte, Denise Keitel, Anna Kaufhold, Kendra K Maass, Kristian W Pajtler, Christof Sohn, Sarah Schott |
Title | 554P KRAS-dependent and independent mechanisms of progressive disease (PD) in colorectal cancer (CRC) patients (pts) with liver metastases (LM) while monitoring on circulating cell free DNA (cfDNA) |
Journal | Annals of Oncology. October 23, 2018. |
Authors | A Kirov Z Mihaylova, V Petrova, T Todorov, D Petkova, A Garev, A Todorova-Georgieva |
Documents | Poster |