PAXgene Blood DNA Kit
全血からのゲノムDNA（最高500 µg）精製
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QIAsymphony PAXgene Blood ccfDNA Kit (192)
カタログ番号 / ID. 768536
Reagent cartridges, accessories and proteinase K vials for 192 preps
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研究用途専用。診断用ではありません。いかなる主張または表示も、疾病の診断、予防、治療のための情報を提供することを目的としてはいません。
このソリューションをお試しになりませんか？
お見積りが必要な場合やプロジェクトについてご相談がある場合は、当社のスペシャリストまでご連絡ください。
特徴
- 1つのシステムで採血および精製を実現
- 簡単な取り扱い
- ワークフローの効率を向上
製品詳細
PAXgene Blood DNAシステムは、採血、安定化、輸送用のパクスジーンDNA採血管（日本ベクトン・ディッキンソン株式会社より入手可能、cat. no. 761115）と、DNA精製用に1本の遠心管で調製できるPAXgene Blood DNA Kitで構成されています。
パフォーマンス
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.
図参照
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).
図参照
操作手順
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).
図参照
アプリケーション
DNAは次のような様々なダウンストリームアプリケーションに使用可能です。
PCRおよびリアルタイム定量PCR
サザンブロット法
SNPジェノタイピング
薬理ゲノミクス研究
SNPディスカバリーとバリデーション
裏付けデータと数値
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).
仕様
| 特徴 | 仕様 |
|---|---|
| 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 |
リソース
MSDS (1)
サイエンティフィック・ポスター (4)
アプリケーション／プロトコール (1)
実験器具関連資料 (1)
キットハンドブック (1)
パンフレット (1)
Application Notes (1)
Certificates of Analysis (1)