Blood STASIS 21-ccfDNA 3mL (200)

Blood STASIS 21-ccfDNA 3mL (200)
Product Code: BS21-CF3-200
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Blood STASIS™ 21 - ccfDNA
For collection of whole blood samples and stabilization of circulating cell-free DNA (ccfDNA) for up to 21 days

The Blood STASIS™ 21-ccfDNA is a plastic, evacuated tube used for collection, anti-coagulation, stabilization, transport, and storage of venous whole blood as well as the preservation of circulating, cell-free DNA (ccfDNA).

  • Room temperature sample transport and storage for up to 21 days
  • Stabilization of circulating cell-free DNA (ccfDNA)
  • Direct draw blood tube for collection, stabilization, transport and storage

    For research use only.  Not for use in diagnostic procedures.

                                    

     
 
Blood STASIS 21-ccfDNA stabilization helps prevent apoptosis of blood cells and degradation of blood genomic DNA.
Whole blood samples were collected in the Blood STASIS 21-ccfDNA tube and K2EDTA tube; then incubated for 21 days at room temperature. Genomic DNA was extracted using MagBio HighPrep Blood & Tissue DNA Kit.  Extracted gDNA were analyzed using 1.2% agarose gel electrophoresis.
 
   
 
Plasma DNA was extracted from 1mL plasma from healthy control using MagBio
CF-Kapture 21 Kit. Four-time points were tested (0, 7,14 and 21 days). Extracted plasma DNA was analyzed using Qubit and qPCR with Alu gene-specific primer sets (115bp; ALU 115) and long (247 bp, ALU 245) amplicons from genomic ALU repeats.  ccfDNA  shows small size distribution around 165bp; ALU 115-qPCR results accurately detect the total quantity of ccfDNA and high molecular weight cellular genomic DNA. Alu247-qPCR results indicate only presence of high molecular weight cellular gDNA  contamination. The ratio ALU247/ALU115 is, therefore, an accurate indication ccfDNA integrity of the sample- T.B. hao in the British Journal of Cancer (2014) 111, 1482-1489). According to results from Hao and al. ccfDNA integrity indices are as follow:
 
 


 


 

        

 * RT- room temperature
** Each Ct value number represent average of 3 data points.

Ten (10)mL whole blood samples from healthy male donor were collected in EDTA, Blood STASIS 21-ccfDNA tube and control blood stabilization tubes; and stored at various temperatures (8°C, 22°C (room temperature), 32°C) for consecutive 21 days. At each time point, 1 mL plasma fraction was separated.  qPCR analysis of SRY gene was used to assess the stability of blood samples and ccfDNA. A control sample with known stability was used to establish a cutoff Ct value for PASS of FAIL sample stability test. Ct value of Control sample=31.00

Blood samples stored in EDTA blood collection tube show lower Ct value, from 30.60-24.01, suggesting gradual lysis of blood cell during incubation as temperature rises and therefore release of genomic DNA (gDNA) and possible contamination of cfDNA with gDNA.

Conversely, all blood samples stored in blood STASIS tube, show slight change in Ct value; 31.10 and 31.99, suggesting a small change in DNA content of the samples, and therefore demonstrating absence of genomic DNA contamination and stability in cfDNA content.

Concluding Remark: Blood STASIS 21-ccfDNA tube allows stabilization of whole blood sample when stored at temperature up to 32°C for 21 consecutive days

                                     

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