DNeasy Blood & Tissue Kit

動物の血液、組織および細胞、酵母、バクテリア、ウイルスからのトータルDNAの精製用

S_1405_RPA_DNY0900
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DNeasy Blood & Tissue Kit (50)

Cat. No. / ID:  69504

50 DNeasy Mini Spin Columns, Proteinase K, Buffers, Collection Tubes (2 ml)
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$224.00
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Kit
DNeasy Blood & Tissue Kit
DNeasy Blood & Tissue QIAcube Kit
Eco-friendlier kit
Column typePlate type
Mini
96-well
Preparations
50
250
DNeasy Blood & Tissue 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。
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For an eco-friendlier alternative to the selected product, look no further.

Features

  • 様々なサンプルタイプに標準化された方法
  • 特殊なサンプルからも高収量で精製
  • 高品質なDNA
  • 様々なスタートサンプルに至適化済みのプロトコール
  • スピンカラムおよびハイスループット用96ウェルフォーマット

Product Details

DNeasy Blood & Tissue Kit は、簡便なスピンカラムおよび96 ウェルプレートフォーマットにおけるシリカベースの迅速で容易なDNA精製を実現します。ほとんどのサンプルはProteinase Kで直接溶解できるため、機械的な破砕の必要はなく、マニュアルでの作業の時間を短縮できます。サンプルタイプ別に至適化されたプロトコールは、ライフサイエンス、ジェノタイピング、獣医学における病原体研究などのアプリケーションに適した再現性の高い高品質なDNAの精製を実現します。DNeasy Blood & Tissue Kitを用いたDNA精製はQIAcube上で自動化できます。

Performance

効率的なDNeasy Blood & Tissue精製法は、動物血液および組織サンプルから収量の高いDNA精製を実現します(表"DNeasy Blood & Tissue Kitsによる動物組織からの一般的なDNA収量"および図"DNA収量")。至適化済みのプロトコールにより、培養細胞、固定化された組織、グラム陽性/陰性菌などと同様に、動物の毛(図" ウマのジェノタイピング")のような一般的ではないサンプルからも高収量で精製できます。酵母、昆虫、毛、その他のサンプルタイプからのDNA精製用に特化されたプロトコールをオンライン上で提供しています。

DNeasy Blood & Tissue Kitによる動物組織からの一般的な収量
由来 DNA(µg)
哺乳動物の血液 100 µl 3~6
鳥類の血液 5 µl 9~40
HeLa細胞 2 x 106 15~25
肝臓 25 mg 10~30
25 mg 15~30
腎臓 25 mg 15~30
脾臓 10 mg 5~30
マウス尾先端部 1.2 cm(先端部) 10~25
ラット尾先端部 0.6 cm(先端部) 20~40
ブタの耳 25 mg 10~30
ウマの毛 10本 2~4
魚のヒレ 20 mg 10~20
魚の卵(サバ) 10 mg 5~10

DNeasy Blood & Tissue Kitsはライフサイエンス、獣医学関係、ジェノタイピング研究の対象である動物種を含む幅広いサンプルタイプからのDNA精製を簡便化できます(図" 高品質なDNA")。精製したDNAはPCRインヒビターを含まず、通常のPCR、マルチプレックスPCR(図" 効率的な16plex PCR")、リアルタイムPCR(図" リアルタイムPCR")で高感度な検出を実現します。DNeasy Blood & Tissue Kitsを用いてトランスジェニックマウスの解析(図"ハイスループット精製")から家畜繁殖プログラム(図" ブタのジェノタイピング")やジェノタイピングによる血統の確認まで信頼できる結果を実現できます。検体数が多い場合には、DNeasy 96 Blood & Tissue Kitを用いて簡単に処理数を増やすことができます(図"ハイスループット精製")。

See figures

Principle

DNeasy Blood & Tissue Kitsは新鮮/凍結動物組織および細胞、血液、バクテリアなどの様々なサンプルからトータルDNA(例、ゲノムDNA、ミトコンドリアDNA、病原体DNA)を迅速に精製できるようデザインされています。

DNeasyメンブレンは、シリカベースメンブレンの結合特性と単純なマイクロスピンテクノロジーまたはQIAGEN 96ウェルプレート遠心システムを組み合わせたものです。DNAは、溶液中の水和分子から水を除去する高濃度のカオトロピック塩の存在下で、DNeasyメンブレンに吸着されます。DNeasy Blood & Tissue精製法におけるバッファー条件は、DNAのシリカゲルメンブレンへの特異的吸着ならびに夾雑物および酵素阻害物質の最適な除去を可能にするようデザインされています。

精製にはフェノールまたはクロロホルム抽出、あるいはアルコール沈殿が不要であり、マニュアルでの作業が最小限で済みます。従って、DNeasy Blood & Tissue Kitsは多数のサンプルの同時並行処理に最適です。ハイスループットアプリケーションには、DNeasy 96 Blood & Tissue Kitを用いて96または192のサンプルの同時並行処理が可能です。

Procedure

簡便なスピンカラムあるいは96ウェルフォーマットによる確実なシリカゲルメンブレンテクノロジーは、迅速で再現性の高いDNA精製を実現し、有機溶媒抽出やアルコール沈殿の必要はありません(フローチャート" DNeasy Miniおよび96精製法")。サンプルはまずproteinase Kを用いて溶解します。緩衝条件を調整してDNA結合の最適な条件を提供し、ライセートをDNeasy Mini Spin ColumnまたはDNeasy 96プレートにロードします。遠心操作中、夾雑物はDNeasyメンブレンを通過しますが、DNAは選択的に結合します。残存する夾雑物および酵素阻害物質は効率的な2回の洗浄ステップで除去されますが、DNAは水またはバッファーで溶出され、即使用可能です。

DNeasy Mini Spin Columnはコレクションチューブにセット済みで、個別に包装されているので使いやすく安全です。DNeasy Mini Spin Columnを用いた精製手順は、QIAcube上で自動化可能です。DNeasy 96 Blood & Tissue KitはQIAGEN 96ウェルプレート遠心システムを用いて96ウェルフォーマットでハイスループット処理できます。

See figures

Applications

DNeasy Blood & Tissue Kitで精製されたDNAは、以下のようなアプリケーションを含むダウンストリームアッセイに最適です。

ライフサイエンス分野の研究
家畜の繁殖
ジェノタイピングによる血統試験
動物感染研究
ルーチン試験

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsPCR, real-time PCR, genotyping
Elution volume100–200 µl
Time per run or per prep20 minutes – 1 hour
Main sample typeBlood, tissue
Format96-well plate, spin column
Sample amount100 µl/25 mg
ProcessingManual
Yield6 µg/30 µg
TechnologySilica technology
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinDNA

Resources

Supplementary Protocols (3)
This protocol is designed for purification of DNA from up to 50 mg of insects, such as drosophila.
This protocol is designed for purification of DNA from up to 5 x 107 yeast cells.
This protocol is designed for purification of DNA from a 200 μl crude lysate.
MSDS (1)
Download Safety Data Sheets for QIAGEN product components.
サイエンティフィック・ポスター (1)
キットハンドブック (3)
For purification of total DNA from animal blood, animal tissue, rodent tails, ear punches, cultured cells, fixed tissue, bacteria, insects

June 2023
For use on QIAcube Connect
Technical Information and Important Notes (1)
Certificates of Analysis (1)
Kit Handbooks (3)
For use on QIAcube Connect
For purification of total DNA from animal blood, animal tissue, rodent tails, ear punches, cultured cells, fixed tissue, bacteria, insects

June 2023

Publications

The unique 16S rRNA genes of piezophiles reflect both phylogeny and adaptation.
Lauro FM; Chastain RA; Blankenship LE; Yayanos AA; Bartlett DH;
Appl Environ Microbiol; 2006; 73 (3):838-45 2006 Dec 8 PMID:17158629
Assays to detect beta-tubulin codon 200 polymorphism in Trichuris trichiura and Ascaris lumbricoides.
Diawara A; Drake LJ; Suswillo RR; Kihara J; Bundy DA; Scott ME; Halpenny C; Stothard JR; Prichard RK;
PLoS Negl Trop Dis; 2009; 3 (3):e397 2009 Mar 24 PMID:19308251
Whole genome amplification for array comparative genomic hybridization using DNA extracted from formalin-fixed, paraffin-embedded histological sections.
Huang J; Pang J; Watanabe T; Ng HK; Ohgaki H;
J Mol Diagn; 2009; 11 (2):109-16 2009 Feb 5 PMID:19197000
Real-time PCR detection of pathogenic microorganisms in roof-harvested rainwater in Southeast Queensland, Australia.
Ahmed W; Huygens F; Goonetilleke A; Gardner T;
Appl Environ Microbiol; 2008; 74 (17):5490-6 2008 Jul 11 PMID:18621865
MicroRNA-137 targets microphthalmia-associated transcription factor in melanoma cell lines.
Bemis LT; Chen R; Amato CM; Classen EH; Robinson SE; Coffey DG; Erickson PF; Shellman YG; Robinson WA;
Cancer Res; 2008; 68 (5):1362-8 2008 Mar 1 PMID:18316599

FAQ

Do you have a protocol for the isolation of genomic DNA from bone?

Yes, we have the following protocols:

  • Isolation of genomic DNA from compact bone using the QIAamp DNA Mini Kit (QA02)
  • Isolation of genomic DNA from compact bone using the DNeasy Blood & Tissue Kit (DY01)
  • Purification of genomic DNA from bones using the QIAamp DNA Micro Kit (QA39)
  • Purification of archive-quality DNA from bone fragments using the Gentra Puregene Tissue Kit (PG38).
FAQ ID -908
Do you have a protocol for purification of total DNA from crude lysates?

Yes, please follow the Supplementary Protocol 'Purification of total DNA from crude lysates using the DNeasy Blood & Tissue Kit' (DY15).

 

FAQ ID -1255
Do you have a protocol for isolation of genomic DNA from saliva and mouthwash?

Yes, we have the following protocols:

  • Isolation of genomic DNA from saliva and mouthwash using the QIAamp DNA Mini Kit; vacuum procedure (QA19v)
  • Isolation of genomic DNA from saliva and mouthwash using the QIAamp DNA Mini Kit; spin procedure (QA19s)
  • Isolation of genomic DNA from saliva using the DNeasy Blood & Tissue Kit; spin procedure (DY07)
FAQ ID -917
When should carrier be used with the QIAamp DNA Mini or the DNeasy Blood & Tissue Kit?

For DNA isolation using the QIAamp DNA Mini, or the DNeasy Blood & Tissue Kit, we recommend using carrier DNA when expected yields are below 10 ng. If possible, carrier DNAs such as poly-dA, poly-dT or poly-dA:dT should be used. Other carrier DNAs such as herring sperm DNA may interfere with subsequent PCR by binding primers nonspecifically.

Please note that poly-dA may interfere with oligo-dT primers, and, in this case, a different carrier DNA should be used. The concentration of carrier DNA should be at least 10 µg/ml. Optimal amounts need to be determined empirically for each application. The size distribution of carrier DNAs is typically in the range of 100 bp to 10 kb.

FAQ ID -100
Is it possible to stop the DNeasy tissue protocol and store the tissue lysates after digesting in buffer ATL and Proteinase K?
After proteinase K digestion, tissue samples can be stored in Buffer ATL for up 6 months at ambient temperature without any reduction in DNA quality.
FAQ ID - 3362
Are there important considerations for plasma generation and urine handling?

It is strongly advised to follow the recommendations for preparing sample material provided in the corresponding Protocol Sheet to ensure reliable results.

Plasma: It is recommended to perform plasma separation immediately after blood collection when using EDTA as anticoagulant to prevent the release of genomic DNA into the plasma fraction.

Urine: Because circulating cell-free DNA in non-stabilized urine samples is rapidly degraded after sample collection due to high nuclease activity, eluates may contain no DNA or exhibit low DNA concentration. Therefore, it is recommended to stabilize urine samples. Even when using stabilized urine, it is recommended to perform a centrifugation step immediately after stabilization to prevent the release of genomic DNA from cells. Alternatively, non-stabilized urine samples can be processed immediately after collection and centrifugation using ATL pretreatment and automated DNA extraction as described in the corresponding Protocol Sheet.

FAQ ID - 3699
Do you have a protocol for Acetyl Cysteine (NALC) treatment of viscous samples?

Yes, please follow either of the User-Developed Protocols:

FAQ ID -913
Do you have a protocol for purification of total DNA from yeast?

Yes, please follow the Supplementary Protocol 'Purification of total DNA from yeast using the DNeasy Blood & Tissue Kit' (DY13).

 

FAQ ID -1253
What is the expected yield of genomic DNA isolated from bacteria with the DNeasy Blood & Tissue or QIAamp DNA Mini Kit?

The expected yield of genomic DNA isolated from bacteria with the DNeasy Blood & Tissue Kit or the QIAamp DNA Mini Kit is approximately 10 ug of DNA per 2x 109 bacterial cells. Note that yields of genomic DNA will vary depending on bacterial strain, quality of the starting material, growing conditions, and the amount of material processed.

 

FAQ ID -632
Do you have a protocol for the isolation of DNA from soft tissues using the TissueLyser?
303 - Can I use my own lysis buffer with the DNeasy Blood & Tissue or QIAamp DNA Mini Kit?

If you have optimized the lysis conditions for a specific sample type, you can lyse the sample in your lysis buffer, and follow the Supplementary Protocol 'Purification of total DNA from crude lysates using the DNeasy Blood & Tissue Kit' (DY15), or the corresponding protocol in the Appendix of the QIAamp DNA Mini Kit and QIAamp DNA Blood Mini Kit Handbook.

 

Can I use your QIAvac 24 Plus for extraction of DNA from tissues using the DNeasy Blood & Tissue Kit?

We do not recommend using a vacuum manifold for DNA extraction of tissues using the DNeasy Blood & Tissue Kit. Viscosity of the lysates varies from sample to sample; therefore, the DNeasy mini spin columns are at great risk of clogging.

FAQ ID -9154
Do you have a protocol for purification of total DNA from insects?

Yes, please follow the Supplementary Protocol 'Purification of total DNA from insects using the DNeasy Blood & Tissue Kit' (DY14).

 

FAQ ID -1254
How can I precipitate genomic DNA using isopropanol?

Alcohol precipitation is commonly used for concentrating, desalting, and recovering nucleic acids. Since less alcohol is required for isopropanol precipitation, this is the preferred method for precipitation of DNA from large volumes. In addition, isopropanol precipitation can be performed at room temperature, which minimizes co-precipitation of salt that interferes with downstream applications.

 

Procedure

  1. Adjust the salt concentration, for example, with sodium acetate (0.3 M, pH 5.2, final concentration) or ammonium acetate (2.0–2.5 M, final concentration).
  2. Add 0.6–0.7 volumes of room-temperature isopropanol to the DNA solution and mix well.
  3. Centrifuge the sample immediately at 10,000–15,000 x g for 15–30 min at 4°C
  4. Carefully decant the supernatant without disturbing the pellet.
  5. Wash the DNA pellet by adding 1–10 ml (depending on the size of the preparation) of room-temperature 70% ethanol. This removes co-precipitated salt and replaces the isopropanol with the more volatile ethanol, making the DNA easier to redissolve.
  6. Centrifuge at 10,000–15,000 x g for 5–15 min at 4°C.
  7. Carefully decant the supernatant without disturbing the pellet.
  8. Air-dry the pellet for 5–20 min (depending on the size of the pellet).
  9. Redissolve the DNA in a suitable buffer.

Tip: Use a buffer with a pH of 7.5–8.0, as DNA does not dissolve easily in acidic buffers. Often distilled water can have an acidic pH. The addition of EDTA protects the DNA from DNase digestion.

Tip: High-molecular-weight DNA, such as genomic DNA, should be redissolved very gently to avoid shearing. If the DNA pellet does not dissolve easily, heat at 55°C for 1–2 h with gentle shaking.

FAQ ID -2953
Do you have a protocol for isolation of genomic DNA from insects?

Yes, we have the following protocols:

  • Isolation of genomic DNA from mosquitoes or other insects using the QIAGEN Genomic tip (QG06).
  • Purification of total DNA from insects using the DNeasy Blood & Tissue Kit (DY14).
FAQ ID -904
How can I improve DNA yields from very tough tissues using the DNeasy Blood & Tissue Kit or the QIAamp DNA Mini Kit?

Efficient DNA isolation requires thorough sample disruption and digestion.

Although the QIAamp and DNeasy procedures requires no mechanical disruption of the tissue sample, the lysis time will be reduced if the sample is ground in liquid nitrogen or mechanically homogenized in advance. For mechanical homogenization, a rotor–stator homogenizer, such as the QIAGEN TissueRuptor, or a bead mill, such as the QIAGEN TissueLyser, can be used.

To improve digestion of tough tissue samples, Proteinase K incubation at 56°C can be performed overnight. DNA yields may be improved by increasing the amount of Proteinase K or by adding additional proteinase K after several hours of digestion.  

FAQ ID -374
Can I use your QIAvac 96 for extraction of DNA using the DNeasy 96 Blood & Tissue Kit?

We do not recommend using a vacuum manifold for DNA extraction with the DNeasy 96 Blood & Tissue Kit because:

  1. Viscosity of the lysates varies from sample to sample. Therefore, the wells of the DNeasy 96 plate are at great risk of clogging.
  2. Use of vacuum manifold would result in incomplete removal of ethanol and salts after washing with Buffer AW2. This will affect the purity and yield of samples.
FAQ ID -9157
What is the shelf-life for QIAGEN Proteinase K (cat. no. 19131, 19133)?

QIAGEN Proteinase K is stable for up to 1 year after delivery when stored at room temperature. To prolong the shelf-life of Proteinase K, storage at 2–8°C is recommended.

FAQ ID - 3447
Do you have a protocol for the isolation of genomic DNA from sperm?

Yes, we have the following protocols:

  • Isolation of genomic DNA from sperm using the QIAamp DNA Mini Kit; protocol 1 (QA03), long procedure
  • Isolation of genomic DNA from sperm using the QIAamp DNA Mini Kit; protocol 2 (QA04), short procedure
  • Purification of total DNA from animal sperm using the DNeasy Blood and Tissue kit; protocol 1 (DY02), long procedure
  • Purification of total DNA from animal sperm using the DNeasy Blood and Tissue kit; protocol 2 (DY03), short procedure
  • Purification of DNA from epithelial cells mixed with sperm cells using the QIAamp DNA Micro Kit (QA40).
FAQ ID -909
What can be used as an alternative to the A260 measurement for quantification of small amounts of RNA and DNA?

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
What dedicated QIAcube Kits are available?
Do you have a protocol for the isolation of viral DNA from stool?
Yes, please follow the User-Developed Protocol 'Isolation of viral DNA from stool using the DNeasy Blood & Tissue Kit' (DY05).
FAQ ID -929
Do you have a protocol for isolation of genomic DNA from saliva using the DNeasy Blood & Tissue Kit?

Yes, please follow the User-Developed Protocol 'Isolation of genomic DNA from saliva using the DNeasy Blood & Tissue Kit; spin procedure' (DY07).

 

FAQ ID -931
Do you have a protocol for purification of DNA from cultured animal cells using the DNeasy 96 Blood & Tissue Kit?

Yes, please follow the Supplementary Protocol 'Purification of DNA from cultured animal cells using the DNeasy 96 Blood & Tissue Kit' (DY12).

 

FAQ ID -934
What is the composition of buffer AE?

The composition of Buffer AE is:

  • 10 mM Tris-Cl
  • 0.5 mM EDTA; pH 9.0.
FAQ ID -730
Do you have data showing effects of sample size on DNA yield and purity using the DNeasy 96 Blood & Tissue kit?
How do I safely inactivate biohazardous flow-through material?

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
3192 - Is QIAGEN Protease compatible with Buffer ATL?
No, QIAGEN Protease is not compatible with Buffer ATL.
Is the quality and size of DNA extracted with the DNeasy Blood & Tissue kit good enough to generate DNA-libraries for next generation sequencing?
The size of DNA obtained with DNeasy Blood & Tissue kit ranges between 100 bp and 50 kb, with 30 kb fragments predominating. The 30 kb fragments are a good starting point for most of the library preparations. Furthermore, the purified DNA is free of protein, nucleases, and other contaminants and inhibitors, and therefore is suitable for NGS.
FAQ ID - 3517