PAXgene Blood DNA Kit

全血からのゲノムDNA（最高500 µg）精製

✓ オンライン注文による24時間年中無休の自動処理システム

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✓ 迅速で信頼性の高い（再）注文

PAXgene Blood DNA Kit (25)

Cat. No. / ID: 761133

Processing tubes and buffers for 25 preparations

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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。

✓ オンライン注文による24時間年中無休の自動処理システム

✓ 知識豊富で専門的な製品&テクニカルサポート

✓ 迅速で信頼性の高い（再）注文

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

PAXgene Blood DNA Systemを用いて精製したDNAの A₂₆₀/A₂₈₀比は1.7～1.9です。断片のサイズは20～200 kbであり、平均は50～150 kbです（図""）。平均DNA収量は、血液サンプル中に存在する有核細胞数によって、150～500 µgの範囲となります。精製したDNAはPCRおよびリアルタイム定量PCRに用いることができます（図""）。

図参照

High quality and high molecular weight of genomic DNA.

Efficient multiplex PCR of 3 mitochondrial genes.

原理

PAXgene Blood DNA Systemは全血サンプルの採取および安定化とそのゲノムDNAの単離を統合および標準化したシステムです。本システムは、採血および安定化に用いるパクスジーンDNA採血管と併用する必要があり、続けてPAXgene Blood DNA Kitを用いてDNA単離を行ないます。

血液凝固を防ぎ、白血球細胞を安定化する試薬を適切に混合および充填したパクスジーンDNA採血管に採血します。血液サンプルは室温で最大14日間チューブ内で保存可能です。チューブ内の血液検体からDNAはPAXgene Blood DNA Kitに添付されている適切なバッファーを用いて単離されます。得られたゲノムDNAは高純度で、様々なダウンストリームアプリケーションに最適です。

操作手順

PAXgene Blood DNAの操作手順はフローチャート" PAXgene Blood DNA製法"に示しています。血液サンプル（8.5 ml）を、保存および輸送用のパクスジーンDNA採血管に採取します。DNA単離は、血液を調製用遠心管（細胞溶解バッファーを既に充填済みで供給）に加え、赤血球と白血球を溶解するために溶液を混和します。細胞核およびミトコンドリアを遠心操作によりペレット化し、洗浄、溶解バッファーに再懸濁します。プロテアーゼとのインキュベーションにより混在するタンパク質を除去します。イソプロパノールにより沈殿したDNAを70%エタノールで洗浄し、乾燥後再懸濁バッファー中に懸濁します。

図参照

PAXgene Blood DNA procedure.

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

PAXgene Blood DNA procedure.

Blood samples (8.5 ml) are collected in PAXgene Blood DNA Tubes, where they may be stored or transported. For DNA isolation, the blood is transferred to processing tubes (supplied already filled with cell lysis buffer), and the solution is mixed to lyse red and white blood cells. Cell nuclei and mitochondria are pelleted by centrifugation, washed, and resuspended in digestion buffer. Protein contaminants are removed by incubation with a protease. DNA is precipitated in isopropanol, washed in 70% ethanol, dried, and resuspended in resuspension buffer.

Specifications

Features	Specifications
Main sample type	Whole blood
Time per run	60 min/8 samples
Elution volume	1 ml
Yield	150–500 µg
Format	Processing tube (salting out method)
Sample amount	8.5 ml
Applications	(RT-)PCR, (RT-)qPCR, NGS, genotyping, microarray analysis
Processing	Manual (centrifugation)

リソース

For collection, storage, and transport of whole blood for genomic DNA isolation

Second edition — innovative tools

Download Safety Data Sheets for QIAGEN product components.

Groelz et al., ASHG 2004

For isolation of genomic DNA from 8.5 ml human whole blood

FAQ

You can find additional information relating to the PreAnalytiX PAXgene products on the PreAnalytiX website .

FAQ ID - 3515

Buffer BG4 consists of 10 mM Tris·Cl, pH 8.5.

FAQ ID - 3501

No. DNA purified with the PAXgene Blood DNA System is free of detectable RNA contamination. In contrast to commercial salting out methods, additional RNase digestion steps are not necessary.

FAQ ID - 3499

See subsequent FAQs for information about storage interval and temperature. Each laboratory should validate shipping procedures.

FAQ ID - 3507

Samples in PAXgene Blood DNA Tubes can be stored at room temperature (18–25°C) for up to 14 days after blood collection.

FAQ ID - 3508

The typical A260/A280 ratio is from 1.7 to 1.9. The DNA molecules are up to 200 kb in size, with fragments of 50–150 kb predominating.

FAQ ID - 3497

Yes, samples in PAXgene Blood DNA Tubes can be stored at 30°C for up to 3 days after blood collection. However, it is not recommended to store blood samples at temperatures higher than 30°C and storage at high temperatures should not exceed 3 days.

FAQ ID - 3509

The typical A₂₆₀/A₂₈₀ ratio is from 1.7 to 1.9.

FAQ ID - 3511

Yield of genomic DNA varies depending on the white blood cell count of the patient blood sample, the extraction method, and the elution volume. However, DNA yield typically ranges between 16–135 µg/ml whole blood. Specifically, DNA yields from magnetic bead-based extraction on the QIAsymphony ranges between 16.9–51.7 µg/ml whole blood, from silica membrane-based extraction on the QIAcube 17.1–45.8 µg/ml whole blood, and from salting-out precipitation 4.9–53.7 µg/ml whole blood.

FAQ ID - 3510

Yes, mitochondrial and genomic DNA are both isolated.

FAQ ID - 3498

No. The volume of Buffer BG1 (lysis buffer) prefilled in the processing tubes, and all the other buffer volumes are optimized for use with blood from one PAXgene Blood DNA Tube.

FAQ ID - 3500

To date, DNA purified from blood samples in PAXgene Blood DNA Tubes has been successfully tested for use in PCR, including multiplex PCR.

FAQ ID - 3512

DNA purified using the PAXgene Blood DNA System should be dissolved in Buffer BG4 and stored at 2–8°C. For long-term storage, DNA can be stored at –20°C but multiple freezing and thawing cycles should be avoided.

FAQ ID - 3502

Yes. After dissolving the nuclei pellet in digestion buffer, Buffer BG3 (step 7), samples can be stored up to 7 days at 2–8°C or for one day at room temperature (18–25°C). The protocol can also be interrupted after adding isopropanol (step 10).

FAQ ID - 3503

So far, we have shown that DNA purified using the PAXgene Blood DNA System can be used in PCR, including long range and multiplex PCR, restriction digestion and Southern analysis, as well as in various SNP detection procedures including,TaqMan®, LightCycler and Masscode technologies.

See trademarks.

FAQ ID - 3506

Do not add bleach or acidic solutions directly to the sample preparation waste. CAUTION: The sample preparation waste contains guanidinium hydrochloride from Buffer BG3, which can form highly reactive compounds when combined with bleach. For more information please consult the appropriate material safety data sheets (MSDS).

FAQ ID - 3505

The system is designed for use only with human blood samples.

FAQ ID - 3504

Always dispose of potentially biohazardous solutions according to your institution’s waste-disposal guidelines. Although the lysis and binding buffers in QIAamp, DNeasy, and RNeasy kits contain chaotropic agents that can inactivate some biohazardous material, local regulations dictate the proper way to dispose of biohazards. DO NOT add bleach or acidic solutions directly to the sample-preparation waste. Guanidine hydrochloride in the sample-preparation waste can form highly reactive compounds when combined with bleach.
Please access our Material Safety Data Sheets (MSDS) online for detailed information on the reagents for each respective kit.

FAQ ID -12