QIAsymphony PAXgene Blood ccfDNA Kit
For purification of ccfDNA from plasma using the QIAsymphony SP
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QIAsymphony PAXgene Blood ccfDNA Kit (192)
Cat no. / ID. 768536
Reagent cartridges, accessories and proteinase K vials for 192 preps
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.
Want to try this solution for the first time?
Get in touch with our specialists if you need a quote or if you would like to discuss your project.
Features
- 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
Product Details
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).
Performance
The fully automated QIAsymphony PAXgene Blood ccfDNA Kit procedure generates ccfDNA yields comparable to the manual QIAamp Circulating Nucleic Acid Kit procedure (see figure High performance, comparable to QIAamp Circulating Nucleic Acid Kit). Since yield is strongly dependent of donor, individual yields may vary. Onboard measures of the QIAsymphony SP minimize cross-contamination.
See figures
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 (see figure PCR results from blood collected in PAXgene Blood ccfDNA Tubes remain stable over storage time). 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 (see figure Isolated ccfDNA is compatible with methylation-specific downstream applications).
See figures
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 (see figure The QIAsymphony SP processes samples containing magnetic particles). 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 (see figure Two protocol lines of the QIAsymphony PAXgene Blood ccfDNA Kit for different research needs).
See figures
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
Supporting data and figures
Isolated ccfDNA is compatible with methylation-specific downstream applications.
Whole blood of 4 healthy donors was collected into either EDTA or PAXgene Blood ccfDNA Tubes. Blood was left at room temperature for up to 7 days. Each day, ccfDNA was isolated from samples, bisulphite converted (Epitect FAST kit, QIAGEN) and analyzed with the GEMINI assay (Clinical Genomics) based on 2 methylated (BCAT1 and IKZF1) and 1 control (ACTB) target. Three PCR replicates per target sequence were analyzed. ccfDNA from blood stored in EDTA tubes generated more false positives than from blood stored in PAXgene Blood ccfDNA Tubes (data courtesy of Clinical Genomics).
Specifications
| Features | Specifications |
|---|---|
| applications | Wide spectrum of research applications, including analysis of cancer tumor DNA and non-invasive prenatal testing |
| format | Magnetic beads |
| inputvolume | 2.4 ml or 4.8 ml plasma |
| throughput | 192 samples per working day and instrument |
| timeperrun | 96 samples of 4.8 ml plasma in 5 h 40 min |
| processing | Manual: plasma centrifugation Automated: QIAsymphony SP |
| shelflife | 6 weeks for an open reagent cartridge |
| elutionvolume | 60 µl for the STA protocol line 60, 100 or 150 µl for the LAF protocol line |
Resources
Safety Data Sheets (1)
Scientific Posters (4)
Application/Protocol Documents (1)
Kit Handbooks (1)
Labware Files and Documents (1)
Brochures & Guides (1)
Application Notes (1)
Certificates of Analysis (1)