DNeasy Blood & Tissue Kit

从动物血液和组织、细胞、酵母、细菌或者病毒中纯化总DNA

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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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 旨在用于分子生物学应用。这些产品不能用于疾病诊断、预防和治疗。
Is sustainability important to you?
For an eco-friendlier alternative to the selected product, look no further.

Features

  • 为各种类型样本提供标准化的纯化方法
  • 即使是特殊类型起始样本也可以获得高产量DNA
  • 高品质DNA
  • 经过优化的操作方法适合处理多种起始样本
  • 可选择离心柱方式或96孔板高通量方式

Product Details

DNeasy Blood & Tissue Kit采用基于硅胶膜技术的离心柱或96孔板方式快速方便的纯化DNA。大多数样本可直接用蛋白酶K消化处理,无需机械破碎,减少了手动操作时间。为特殊样本提供优化的纯化方法,获得高重复性、高纯度的DNA,用于生物研究、基因型分析和兽医病理研究。DNA纯化流程可以使用DNeasy Blood & Tissue Kit,在QIAcube全自动核酸纯化仪上自动化进行。

Performance

高效的DNeasy Blood & Tissue操作流程能够从动物血液和组织样本中纯化大量的总DNA(参见下表和" DNA yields")。优化的实验方案可确保非标准样本的高产量,如动物毛发(参见" Genotyping of horses")和培养细胞、固定的组织,或者革兰氏阳性和阴性细菌。独特的在线实验方案参考文库,可从酵母、昆虫、毛发和其他类型样本中纯化DNA。

使用DNeasy Blood & Tissue Kits从动物组织中纯化的DNA产量
样本来源 样本量 DNA(µg)
哺乳动物血液 100 µl 3–6
鸟类血液 5 µl 9–40
HeLa细胞 2 x 106 15–25
肝脏 25 mg 10–30
25 mg 15–30
肾脏 25 mg 15–30
脾脏 10 mg 5–30
小鼠尾 1.2 cm(尾部) 10–25
大鼠尾 0.6 cm(尾部) 20–40
猪耳 25 mg 10–30
马毛 10根毛发 2–4
鱼鳍 20 mg 10–20
鱼卵(鲭鱼) 10 mg 5–10

DNeasy Blood & Tissue Kits简化了从多种样本类型中纯化DNA的流程,包括在生命科学、兽医学和基因分型研究应用中常见的动物种类(参见" High-quality DNA")。纯化得到的DNA不含PCR抑制剂,能在标准多重(参见" Efficient 16plex PCR")real-time PCR(参见" Real-time PCR")中进行灵敏的检测。DNeasy Blood & Tissue Kits确保了可靠的实验结果,从转基因小鼠的实验室分析(参见" High-throughput purification")到家畜的育种项目(参见" Genotyping of pigs")和种系基因分型研究。可以用DNeasy 96 Blood & Tissue Kit方便地扩大常规的检测应用的规模。

See figures

Principle

DNeasy Blood & Tissue Kit适用于从不同的样本中,包括新鲜或冷冻的动物组织和细胞、血液或者细菌,快速纯化总DNA(如,基因组、线粒体和病原体等)。

DNeasy膜整合了硅胶膜的结合特性,经简单显微离心技术或者QIAGEN 96-Well-Plate Centrifugation System处理即可。在高浓度离液盐的环境中,DNA吸附在DNeasy膜上,可以除去溶液中水合分子的水分。DNeasy Blood & Tissue操作流程中设计的缓冲液条件,旨在使DNA特异性吸附到硅胶膜上,并且优化去除污染物和酶抑制剂。

纯化过程不需要苯酚或氯仿抽提或乙醇沉淀,仅涉及少量手工步骤。这使得DNeasy Blood & Tissue Kits非常适合同时处理多个样本。对于高通量的应用,DNeasy 96 Blood & Tissue Kit能够同时处理96或192个样本。

Procedure

可靠的硅胶膜技术,利用便捷的离心柱或96孔板形式,确保了快速可重复的DNA纯化,不再需要有机抽提和乙醇沉淀(参见" DNeasy Mini and 96 procedures")。样本首先用蛋白酶K裂解。调整缓冲条件,以提供最佳的DNA结合条件并将裂解物上样到DNeasy Mini离心柱或DNeasy 96孔板上。在离心过程中,DNA选择性地结合到DNeasy膜上,污染物通过。余下的污染物和酶抑制剂将在两个高效的洗涤步骤中被去除。然后,用水或缓冲液将DNA洗脱,待用。  

DNeasy Mini离心柱预装在收集管中,并且单独封存,确保方便和安全。使用DNeasy Mini离心柱的纯化过程可在QIAcube全自动核酸纯化仪上自动进行。DNeasy 96 Blood & Tissue Kit可使用QIAGEN 96-Well-Plate Centrifugation System以96孔板形式进行高通量处理。

See figures

Applications

DNeasy Blood & Tissue Kits提供高品质DNA,可直接用于多种下游分析,包括:

  • 生命科学研究
  • 家畜育种
  • 种系基因分型研究
  • 兽医病理分析研究
  • 常规的应用检测

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

补充实验方案 (4)
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.
This protocol is designed for purification of DNA from up to 50 mg of insects, such as drosophila.
安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
试剂盒操作手册 (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
Supplementary Protocols (4)
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.

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
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