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QIAsymphony PAXgene Blood ccfDNA Kit

For purification of ccfDNA from plasma using the QIAsymphony SP

  • Optimized chemistry and input volumes for PAXgene Blood ccfDNA Tubes
  • Separate protocol lines for small (STA) and large (LAF) ccfDNA fragments
  • Prefilled reagent cartrdiges with bar code for safety and ease of use
  • Standardized processing for high reproducibility

The PAXgene Blood ccfDNA System consists of a blood collection tube (PAXgene Blood ccfDNA Tube) and a ccfDNA purification kit for the QIAsymphony (QIAsymphony PAXgene Blood ccfDNA Kit).

Артикул: 768536
QIAsymphony PAXgene Blood ccfDNA Kit (192)

Reagent cartridges, accessories and proteinase K vials for 192 preps


Performance

The fully automated QIAsymphony PAXgene Blood ccfDNA Kit procedure generates ccfDNA yields comparable to the manual QIAamp Circulating Nucleic Acid Kit procedure. Since yield is strongly dependent of donor, individual yields may vary. Onboard measures of the QIAsymphony SP minimize cross-contamination.

Principle

Apoptosis and lysis of white blood cells in unstabilized whole blood releases genomic DNA into plasma, which dilutes naturally occurring ccfDNA. PAXgene Blood ccfDNA Tubes contain an additive that prevents blood coagulation and immediately stabilizes blood cells via a non-crosslinking stabilization solution. This impedes the release of intracellular DNA and maintains constant ccfDNA levels in plasma. When used with PAXgene Blood ccfDNA Tubes, the proven magnetic particle technology of the QIAsymphony PAXgene Blood ccfDNA Kit provides a rapid, automated procedure to isolate ccfDNA of high quality and purity.

Procedure

The workflow begins with a centrifugation step that separates plasma from the cellular fraction of the whole blood collected into PAXgene Blood ccfDNA Tubes. The plasma is then carefully transferred into a second tube. A second centrifugation step can be carried out optionally to remove any traces of buffy coat. The pure plasma is then transferred to a third tube which is loaded into the QIAsymphony SP.

In the QIAsymphony SP, plasma proteins are digested by proteinase K while ccfDNA binds to the surface of magnetic beads. Depending on the chosen protocol line, predominantly small ccfDNA fragments (STA protocol line) or small and large ccfDNA fragments (LAF protocol line) are isolated. Three wash steps ensure contaminant removal. Finally, ccfDNA is eluted from the magnetic particles and is ready for use in downstream applications.

Applications

ccfDNA purified using the QIAsymphony PAXgene Blood ccfDNA Kit and the QIAsymphony SP is ready for use in a wide range of downstream applications, including:

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

Features
Specifications
Applications Wide spectrum of research applications, including analysis of cancer tumor DNA and non-invasive prenatal testing
Elution volume 60 µl for the STA protocol line 60, 100 or 150 µl for the LAF protocol line
Format Magnetic beads
Input volume 2.4 ml or 4.8 ml plasma
Processing Manual: plasma centrifugation Automated: QIAsymphony SP
Shelf life 12 weeks for an open reagent cartridge
Throughput 192 samples per working day and instrument
Time per run 96 samples of 4.8 ml plasma in 5 h 40 min