PAXgene Blood DNA Kit
Whole blood에서 genomic DNA를 500 ug까지 분리 정제할 수 있습니다
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특징
- 하나의 system으로 혈액 회수와 정제가 가능함
- 쉬운 사용방법
- Enhanced workflow efficiency
제품 세부 정보
The PAXgene Blood DNA system consists of PAXgene Blood DNA Tubes, for blood collection and stabilization, and the PAXgene Blood DNA Kit, for DNA purification in a single-tube procedure.
성능
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.
그림 참조
원리
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).
그림 참조
절차
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).
그림 참조
응용 분야
The DNA can be used in a wide range of downstream applications, including:
PCR and quantitative, real-time PCR
Southern blotting
SNP genotyping
Pharmacogenomic studies
SNP discovery and validation
지원되는 데이터 및 수치
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).
사양
| 특징 | 사양 |
|---|---|
| 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 |
리소스
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)