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PAXgene Blood ccfDNA Tubes (100)

For collection of whole blood samples and stabilization of ccfDNA

 

  • Singe-tube collection, stabilization, transport and storage
  • Immediate stabilization of circulating, cell-free DNA (ccfDNA)
  • Sample transport and storage at room temperature for up to 7 days
  • Standardized preanalytical processing of samples

The PAXgene Blood ccfDNA Tube is a plastic, closed, evacuated tube for the collection, anticoagulation, transport and storage of human whole blood samples and the stabilization of ccfDNA. When the tube is used in conjunction with the QIAsymphony PAXgene Blood ccfDNA Kit or the QIAGEN QIAamp Circulating Nucleic Acid Kit, high-quality ccfDNA can be isolated for use in downstream analytical assays.
Cat No./ID: 768115
PAXgene Blood ccfDNA Tubes (100)
€907.00
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100 blood collection tubes (10 ml). To be used in conjunction with QIAGEN QIAamp Circulating Nucleic Acid Kit (50) or QIAsymphony PAXgene Blood ccfDNA Kit (192)
For Research Use Only. Not for use in diagnostics procedures. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease.
0
PAXgene Blood ccfDNA stabilization reagent helps prevent release of gDNA into plasma.

Whole blood was stored in EDTA tubes or PAXgene Blood ccfDNA Tubes. ccfDNA was extracted from the plasma immediately following blood collection (t0) or after 6 days storage (t6) at room temperature. Eluate (1 µl) was analyzed using the Agilent High Sensitivity DNA Kit. Plasma from EDTA tubes showed an increase in apoptotic gDNA fragments, whereas plasma from PAXgene Blood ccfDNA Tubes showed a ccfDNA profile comparable to day 0.

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PAXgene Blood ccfDNA stabilization reagent helps prevent hemolysis, allowing clear distinction between plasma and cell fractions.

Whole blood was stored in EDTA tubes or PAXgene Blood ccfDNA Tubes. Hemolysis was determined by measuring the absorption of free hemoglobin in plasma from 6 subjects at 414 nm. Mean absorption and standard deviation are shown. Amount of hemolysis increased with each additional storage time point in the EDTA tubes, but remained relatively constant in the PAXgene Blood ccfDNA additive.

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PAXgene Blood ccfDNA stabilization helps prevent apoptosis and fragmentation of gDNA in whole blood.

Whole blood from 2 subjects was stored in PAXgene Blood ccfDNA Tubes and EDTA tubes. Genomic DNA was extracted from samples using the QIAamp Blood Mini Kit immediately after blood collection and after 10 days of storage. Agarose gel electrophoresis was used for size separation of 400 ng extracted gDNA. Fragmentation was evident in the gDNA extracted from whole blood stored in EDTA tubes, but practically absent from the samples stored in PAXgene Blood ccfDNA Tubes.

3
PAXgene Blood ccfDNA stabilization helps prevent release of gDNA into plasma during storage and transport.

Whole blood from 8 subjects was collected into EDTA tubes (unstabilized) and PAXgene Blood ccfDNA Tubes (stabilized). Samples were stored for 1 day at 35°C (t1, 35°C), for 6 days at room temperature (t6) or to simulate transport, continually inverted for 5 hours and then stored horizontally for 6 days at room temperature (t6sim). ccfDNA was extracted from plasma using the QIAsymphony SP Protocol for the PAXgene Blood ccfDNA System and ccfDNA yield was quantified by real-time PCR (18S rDNA gene, 66 bp, 180 bp and 500 bp amplicons). Shown is the average ratio of copy numbers at testing time (tx) to copy numbers immediately after blood draw (t0), plus standard deviation. The ratio remained close to 1 in all treatments of the PAXgene Blood ccfDNA stabilized samples, indicating the 18S rDNA copy number did not change over time. A significant increase in 18S rDNA copy number was observed in the EDTA samples after storage, indicating release of gDNA into the plasma.

4
PAXgene Blood ccfDNA stabilization minimizes hemolysis compared to alternative solutions.

Whole blood from 20 subjects was collected into PAXgene Blood ccfDNA Tubes, EDTA tubes and blood collection tubes designed for ccfDNA stabilization from 2 other suppliers. Plasma was separated directly after blood draw, after 3 days and after 7 days storage at room temperature (15–25°C). Hemolysis was assessed by measuring plasma free hemoglobin (pfHb) using the Catachem Plasma Free Hemoglobin (PFH) Kit on the Beckman Counter DxC analyzer. Increase in sample hemolysis during sample storage at room temperature was minimized in the PAXgene Blood ccfDNA Tubes compared to the other blood collection tubes with different stabilization solutions. This indicates red blood cell lysis and decreased sample integrity in the alternative blood collection tubes.

Performance
PAXgene Blood ccfDNA Tubes simplify the collection and processing of whole blood for subsequent purification of  circulating, cell-free DNA (ccfDNA). The tube additive stabilizes blood cells (see "PAXgene Blood ccfDNA stabilization helps prevent hemolysis allowing distinct separation of plasma" and "PAXgene Blood ccfDNA stabilization minimizes hemolysis compared to alternative solutions") and blocks apoptosis (see "PAXgene Blood ccfDNA stabilization helps prevent apoptosis and fragmentation of gDNA").  Consequently, using PAXgene Blood ccfDNA Tubes for whole blood collection, storage and transport helps prevent the release of intracellular DNA into plasma (see "PAXgene Blood ccfDNA stabilization helps prevent release of gDNA into plasma"). ccfDNA levels are stable for up to 7 days at room temperature (15–25°C) or up to 24 hours at 35°C, so blood samples can be transported and stored at room temperature to process plasma later (see "PAXgene Blood ccfDNA stabilization prevents release of gDNA into plasma during storage and transport").

Plasma generated from whole blood collected into PAXgene Blood ccfDNA Tubes and processed with the QIAsymphony PAXgene Blood ccfDNA Kit or the QIAGEN QIAamp Circulating Nucleic Acid Kit, yields high-quality ccfDNA for downstream analytical assays, including but not limited to PCR-based assays and next-generation sequencing (NGS).
Principle
The PAXgene Blood ccfDNA Tube is a plastic, closed, evacuated tube for the collection, anticoagulation, transport and storage of human whole blood samples and the stabilization of ccfDNA. The tube contains an additive that anticoagulates blood and stabilizes blood cells via a non-crosslinking stabilization solution. This helps prevent the release of intracellular DNA into the plasma and maintains constant ccfDNA levels during sample storage before processing.
Procedure
The PAXgene Blood ccfDNA Tube allows collection of 10 ml whole blood into a closed, evacuated system. Using standard venipuncture technique, blood is collected into an evacuated tube that contains 1.5 ml cell stabilization additive, resulting in a ratio of 0.15 ml additive per 1 ml blood. Isolation of ccfDNA from plasma is carried out using the QIAsymphony PAXgene Blood ccfDNA Kit or the QIAamp Circulating Nucleic Acid Kit.
Applications

ccfDNA purified from plasma generated from whole blood collected into PAXgene Blood ccfDNA Tubes can be used in a wide range of downstream applications, including:

  • PCR, including digital, multiplex and quantitative real-time PCR
  • Pharmacogenomic studies
  • SNP genotyping
  • Next-generation sequencing

 

Features
Specifications
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Handbooks & Protocols (1)
For collection of whole blood and stabilization of ccfDNA
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Kit Handbooks (1)
For collection of whole blood and stabilization of ccfDNA
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