PAXgene Blood DNA Kit
从全血样本中纯化至多500 μg基因组DNA
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QIAsymphony PAXgene Blood ccfDNA Kit (192)
目录编号 / ID. 768536
Reagent cartridges, accessories and proteinase K vials for 192 preps
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仅作研究用途。不用于诊断程序。任何表述或表示都并非旨在提供诊断、预防或治疗疾病的信息。
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特点
- 同一体系中进行血液收集和纯化
- 易于操作
- 提高工作流程的效率
产品详情
PAXgene Blood DNA系统包括用于血液采集和稳定的PAXgene Blood DNA Tubes和用于单管纯化DNA的PAXgene Blood DNA Kit。
绩效
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.
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原理
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).
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程序
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).
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应用
纯化的DNA可用于多种下游应用，包括：
PCR和定量real-time PCR
Southern印迹
SNP基因分型
药物基因组研究
SNP发现和鉴定
辅助数据和图表
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).
规格
| 特点 | 规格 |
|---|---|
| applications | PCR,qPCR,real-timePCR,blotting,genotyping |
| format | Processingtube |
| inputvolume | Wholeblood |
| throughput | 8.5ml |
| timeperrun | 96 samples of 4.8 ml plasma in 5 h 40 min |
| processing | Manual: plasma centrifugation Automated: QIAsymphony SP |
| shelflife | 150–500µg |
| elutionvolume | 1ml |
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