QIAamp RNA Blood Mini Kit – RNA Extraction from Blood

新鮮血液からの細胞性RNA分離用

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✓ 迅速で信頼性の高い（再）注文

QIAamp RNA Blood Mini Kit (50)

Cat. No. / ID: 52304

For 50 RNA preps: 50 QIAamp Mini Spin Columns, 50 QIAshredder Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free reagents and buffers

KitBuffer

QIAamp RNA Blood Mini Kit

Buffer AW1

Inquire

QIAamp RNA Blood Mini Kitは分子生物学的アプリケーション用であり、疾病の診断、予防、あるいは治療に使用することはできません。

✓ 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

高品質で即使用可能なRNAの迅速な精製
有機溶媒抽出あるいはアルコール沈殿が不要
一定した高い収量
夾雑物および酵素阻害物質を完全除去

製品詳細

QIAamp RNA Blood Mini Kitは、クエン酸、ヘパリン、EDTAのような一般的な抗凝固剤で処理した新鮮なヒト全血（最高1.5 ml）からシリカメンブレンをベースにした精製法で細胞RNAを分離します。QIAshredder Spin Columnを用いたホモジナイゼーション後、迅速なスピンカラム法によりRNAを簡単に精製します。精製はQIAcube上で完全自動化が可能です。

パフォーマンス

QIAamp調製法は、完全にRNase、夾雑物および酵素阻害物質を除去するので、全てのダウンストリームアプリケーションに最適な高品質RNAが得られます（図 " ノーザンブロット解析用の高品質RNA " および " RT-PCR解析"）。

QIAamp RNA Blood Mini Kit を用いれば、DNA のコンタミを最小限に抑えた最高品質のRNAが調製できます。しかしどのようなRNA 精製法を用いても、わずかなDNA コンタミは避けられません。DNA に非常に高感度なRNAアプリケーションの場合には、残存するDNAの完全な除去が必要となります。このような場合には、QIAamp RNA調製中に簡便なカラム上で、サンプルのDNase 処理が行なえるQIAGEN RNase-Free DNase Set のご使用をお薦めします。

図参照

High-quality RNA for northern analysis.

Reliable RT-PCR analysis.

原理

QIAamp RNA Blood Mini Kit を使用すれば、シリカメンブレンテクノロジーによって細胞RNAを精製することができます。フェノール／クロロホルム抽出は不要です。RNAは選択的にQIAampシリカゲルメンブレンに結合し、夾雑物は洗い流されます。2価の陽イオンおよびタンパク質などのPCR阻害物質は2回の効率的な洗浄ステップにより完全に除去され、残った高純度のRNAが水あるいはキットに添付のバッファーで溶出されます。

QIAampテクノロジーによって、新鮮血液などのサンプルから細胞性トータルRNAを精製し、RT-PCRおよびブロッティング操作ですぐに使用することができます。QIAampによるサンプル調製テクノロジーは、ライセンス保証されています。

操作手順

QIAamp RNA Blood Mini Kit は迅速なスピンカラム法により、血液からのRNA精製を容易にします（図 ""）。赤血球を選択的に溶解し、白血球を遠心操作で収集します。その後、RNaseを急速に不活化する高度な変性の条件を使用して、白血球を溶解します。QIAshredder Spin Columnによるホモジナイゼーションの後、サンプルをQIAamp Spin Column にアプライします。トータルRNAがQIAampメンブレンに結合し、夾雑物が洗浄除去され、高純度なRNA が、30～100 µlのRNaseフリーの水（本キットに添付）で溶出されて、全てのダウンストリームアプリケーションで即使用可能となります。

図参照

QIAamp RNA Blood Mini procedure.

アプリケーション

QIAamp RNA Blood Mini Kit は、1.5 ml までのクエン酸塩、ヘパリン、EDTA 等の一般的抗凝血剤入りの新鮮なヒト全血から細胞性RNA を精製するキットです。組織サンプルからは、トータルRNA を本キットで精製することが可能です。

裏付けデータと数値

QIAamp RNA Blood Mini procedure.

Specifications

Features	Specifications
Applications	PCR, real-time PCR, microarray
Elution volume	30–100 µl
Main sample type	Whole blood, tissue
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein	Cellular RNA
Sample amount	50–1.5 ml
Format	Spin columns
Processing	Manual (centrifugation)
Time per run or per prep	<1 hour
Technology	Silica technology
Yield	1–5 µg

リソース

For total RNA purification from human whole blood

Poster for download

Download Safety Data Sheets for QIAGEN product components.

For total RNA purification from human whole blood

Poster for download

Publications

Preanalytical mRNA stabilization of whole bone marrow samples.

Langebrake C; Günther K; Lauber J; Reinhardt D;

Clin Chem; 2007; 53 (4):587-93 2007 Feb 8 PMID:17289802

Histone acetylation dependent allelic expression imbalance of BAPX1 in patients with the oculo-auriculo-vertebral spectrum.

Fischer S; Lüdecke HJ; Wieczorek D; Böhringer S; Gillessen-Kaesbach G; Horsthemke B;

Hum Mol Genet; 2006; 15 (4):581-7 2006 Jan 11 PMID:16407370

Molecular analysis of the GNAS1 gene for the correct diagnosis of Albright hereditary osteodystrophy and pseudohypoparathyroidism.

De Sanctis L; Romagnolo D; Olivero M; Buzi F; Maghnie M; Scirè G; Crino A; Baroncelli GI; Salerno M; Di Maio S; Cappa M; Grosso S; Rigon F; Lala R; De Sanctis C; Dianzani I;

Pediatr Res; 2003; 53 (5):749-55 2003 Mar 5 PMID:12621129

Perforin-dependent brain-infiltrating cytotoxic CD8+ T lymphocytes mediate experimental cerebral malaria pathogenesis.

Nitcheu J; Bonduelle O; Combadiere C; Tefit M; Seilhean D; Mazier D; Combadiere B;

J Immunol; 2003; 170 (4):2221-8 2003 Feb 15 PMID:12574396

FAQ

One milliliter of healthy human blood consists of cell types in approximately the following numbers:

Leucocytes (function: immune response) 4–7 x 106 cells
Thrombocytes (function: wound closing) 3–4 x 108 cells
Erythrocytes (function O2 and CO2 transport) 5 x 109 cells

FAQ ID -2951

Several kit options are available for this application. We recommend using the PAXgene Blood RNA System, which enables the collection, stabilization and transportation of 2.5 ml human whole blood samples, and subsequent rapid and efficient isolation of cellular RNA.

Other products for the isolation of RNA from whole human blood are the QIAamp RNA Blood Mini Kit and the RNeasy Midi Kit for processing up to 1.5 ml and 10 ml human whole blood, respectively.

FAQ ID -304

The exact composition of Buffer RLT is confidential. This buffer is a proprietary component of RNeasy Kits. Buffer RLT contains a high concentration of guanidine isothiocycanate, which supports the binding of RNA to the silica membrane. Buffer RLT can be purchased separately (cat. no. 79216)

Note: note that ß-mercaptoethanol should be added to Buffer RLT before use to effectively inactivate RNAses in the lysate (10 µl ß-Mercaptoethanol per 1 ml Buffer RLT).

FAQ ID -2793

QIAGEN Proteinase K is a subtilisin-type protease, which cleaves at the carboxyl side of hydrophobic, aliphatic and aromatic amino acids. It is particularly suitable for short digestion times. It possesses a high specific activity over a wide range of temperatures and pH values with substantially increased activity at higher temperature. Soluble calcium is not essential for enzymatic activity. This means that EDTA, which is used to inhibit Mg2+-dependent enzymes such as nucleases, will not inhibit Proteinase K activity.

QIAGEN Protease is a broad-specificity Serine protease with high activity, cleaving preferentially at neutral and acidic residues. It is an economical alternative to Proteinase K for isolation of native DNA and RNA from a variety of samples.

FAQ ID -761

Yes. The RNeasy MinElute Cleanup Kit has been developed specifically for cleaning up and concentrating RNA samples. You can also follow the protocols for RNA cleanup in the RNeasy Mini and RNeasy Midi/Maxi Handbook.

FAQ ID -286

Yes. Even though buffer RDD in the RNase-Free DNase Set is optimized for on-column DNase digestion, the buffer is also well-suited for efficient DNase digestion in solution. Please note that the reaction must be cleaned up after the off-column DNase digest to remove the enzyme and buffer RDD, which will interfere with subsequent RT reactions.

A protocol for in-solution DNase digestion using the RNase-Free DNase Set can be found in Appendix C of the RNeasy MinElute Cleanup Handbook. For subsequent RNA Cleanup, use either the RNeasy MinElute Cleanup Kit, or follow the instructions for RNA Cleanup in the RNeasy Mini Handbook.

FAQ ID -619

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728

According to an Interview with Professor Dennis Lo published in QIAGEN News Molecular Diagnostics, Issue No. 5, 2002, healthy individuals have about 500-1000 genome equivalents (DNA) per ml serum/plasma.

For free-circulating DNA in plasma, the concentration can range from 1–100 ng/ml in healthy individuals.

FAQ ID -635

For RNA isolated on the BioRobot EZ1 and BioRobot M48:

The RNA can be directly applied to the Agilent Bioanalyzer, since it is being denatured in the final protocol steps of these isolation procedures.

For RNA prepared with all other QIAGEN RNA Isolation Products:

We recommend to denature the samples in a water bath for 2 min at 70°C, and then place them directly on ice prior to loading them onto the Agilent Bioanalyzer.

FAQ ID -528

The A₂₆₀/ A₂₈₀ ratio is influenced considerably by pH. Since water is not buffered, the pH and the resulting A₂₆₀/A₂₈₀ ratio can vary greatly. Lower pH results in a lower A₂₆₀/ A₂₈₀ ratio and a reduced sensitivity to protein contamination*.

For accurate values, we recommend measuring absorbance in 10 mM Tris·Cl, pH 7.5. Always be sure to calibrate the spectrophotometer with the same solution. Please see the Appendix sections in the RNeasy handbooks for additional information.

* Wilfinger, W.W., Mackey, M., and Chomczynski, P. (1997) Effect of pH and ionic strength on the spectrophotometric assessment of nucleic acid purity. BioTechniques 22, 474.

FAQ ID -97

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

The concentration of RNA isolated with RNeasy Kits can be determined by measuring the absorbance at 260 nm (A260) in a spectrophotometer. Absorbance readings should be greater than 0.15 to ensure significance. An absorbance of 1 unit at 260 nm corresponds to 40 µg of RNA per ml (A260 = 1 = 40 µg/ml). This relationship is valid for measurements in water. Therefore, dilute RNA in water to quantify it spectrophotometrically.

An example of the calculations involved in RNA quantification is shown below. Use the buffer in which the RNA is diluted to zero the spectrophotometer:

Volume of RNA sample = 100 µl
Dilution = 10 µl of RNA sample + 490 µl distilled water (1/50 dilution)
Absorbance of diluted sample measured in a 1 ml cuvette (RNase-free): A260 = 0.23
Concentration of original RNA sample = 40 x A260 x dilution factor = 40 x 0.23 x 50
RNA concentration: 460 µg/ml
Total yield = concentration x volume of sample (ml) = 460 µg/ml x 0.1 ml
RNA yield: 46 µg

For additional information on RNA quantitation and handling, see the Appendix section in the RNeasy Mini Handbook.

FAQ ID -32