DNeasy Blood & Tissue Kits

For spin-column or 96-well extraction of total DNA from animal blood and tissues and from cells, yeast, bacteria, or viruses

Features

  • Standardized method for a variety of sample types
  • High yields even from specialized samples
  • High-quality DNA
  • Optimized protocols for a range of starting materials
  • Spin-column and 96-well high-throughput formats
DNeasy Blood & Tissue Kit (50)

Cat. No. / ID: 69504

50 DNeasy Mini Spin Columns, Proteinase K, Buffers, Collection Tubes (2 ml)
Column typePlate type
Mini
96-well
Preparations
50
250
The DNeasy Blood & Tissue Kits is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

DNeasy Blood & Tissue Kits provide fast and easy silica-based DNA extraction without phenol or chloroform in convenient spin-column and 96-well-plate formats. Most samples can be directly lysed with proteinase K, eliminating the need for mechanical disruption and reducing hands-on time. Optimized protocols for specific sample types provide reproducible extraction of high-quality DNA for life science, genotyping, and veterinary pathogen research applications. Extraction of DNA using the DNeasy Blood & Tissue Kit can be automated on the QIAcube Connect.

Performance

The efficient DNeasy Blood & Tissue procedure enables high yields of total DNA from animal blood and tissue samples (see table "Typical DNA yields from animal tissues using DNeasy Blood & Tissue Kits" and figure "DNA yields"). Optimized protocols ensure high yields from nonstandard samples, such as animal hair (see figure " Genotyping of horses"), as well as cultured cells, fixed tissues, or Gram-positive and -negative bacteria. Specialized online protocols are available for DNA purification from yeast, insects, hair, and other sample types.

Typical yields from animal tissues using DNeasy Blood & Tissue Kits
SourceAmount DNA(µg)
Mammalian blood 100 µl 3–6
Bird blood 5 µl 9–40
HeLa cells 2 x 106 15–25
Liver 25 mg 10–30
Brain 25 mg 15–30
Kidney 25 mg 15–30
Spleen 10 mg 5–30
Mouse tail 1.2 cm (tip) 10–25
Rat tail 0.6 cm (tip) 20–40
Pig ear 25 mg 10–30
Horse hair 10 hairs 2–4
Fish fin 20 mg 10–20
Fish spawn (mackerel) 10 mg 5–10

DNeasy Blood & Tissue Kits simplify purification of DNA from a wide range of sample types, including animal species commonly encountered in life science, veterinary, and genotyping applications (see figure " High-quality DNA"). Purified DNA is free from PCR inhibitors, enabling sensitive detection in standard, multiplex (see figure " Efficient 16plex PCR"), and real-time PCR (see figure " Real-time PCR"). DNeasy Blood & Tissue Kits provide reliable results — from laboratory analysis of transgenic mice (see figure "High-throughput purification") to livestock breeding programs (see figure " Genotyping of pigs") and pedigree genotyping. Routine testing applications can be easily scaled up using the DNeasy 96 Blood & Tissue Kit (see figure "High-throughput purification").

See figures

Principle

DNeasy Blood & Tissue Kits are designed for rapid purification of total DNA (e.g., genomic, mitochondrial, and pathogen) from a variety of sample sources including fresh or frozen animal tissues and cells, blood, or bacteria.

The DNeasy membrane combines the binding properties of a silica-based membrane with simple microspin technology or with the QIAGEN 96-Well-Plate Centrifugation System. DNA adsorbs to the DNeasy membrane in the presence of high concentrations of chaotropic salt, which remove water from hydrated molecules in solution. Buffer conditions in DNeasy Blood & Tissue procedures are designed to enable specific adsorption of DNA to the silica membrane and optimal removal of contaminants and enzyme inhibitors.

Purification requires no phenol or chloroform extraction or alcohol precipitation, and involves minimal handling. This makes DNeasy Blood & Tissue Kits highly suited for simultaneous processing of multiple samples. For higher-throughput applications, the DNeasy 96 Blood & Tissue Kit enables simultaneous processing of 96 or 192 samples.

Procedure

Reliable silica-membrane technology, in convenient spin-column or 96-well formats, ensures fast and reproducible DNA purification, eliminating the need for organic extraction and alcohol precipitation (see flowchart " DNeasy Mini and 96 procedures"). Samples are first lysed using proteinase K. Buffering conditions are adjusted to provide optimal DNA-binding conditions and the lysate is loaded onto the DNeasy Mini spin column or the DNeasy 96 plate. During centrifugation, DNA is selectively bound to the DNeasy membrane as contaminants pass through. Remaining contaminants and enzyme inhibitors are removed in two efficient wash steps and DNA is then eluted in water or buffer, ready for use.

DNeasy Mini spin columns are prepackaged in collection tubes and individually sealed, providing convenience and safety. The purification procedure using DNeasy Mini spin columns can be automated on the QIAcube. The DNeasy 96 Blood & Tissue Kit provides high-throughput processing in a 96-well format using the QIAGEN 96-Well-Plate Centrifugation System.

See figures

Applications

DNeasy Blood & Tissue Kits provide high-quality DNA, ready to use in all downstream assays, including applications in:

Life science research
Livestock breeding
Pedigree genotyping
Veterinary pathogen research
Routine applied testing

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 (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 up to 50 mg of insects, such as drosophila.
This protocol is designed for purification of DNA from a 200 μl crude lysate.
Brochures & Guides (1)
Kit Handbooks (1)
For purification of total DNA from animal blood, animal tissue, rodent tails, ear punches, cultured cells, fixed tissue, bacteria, insects

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 or citrate 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.

 

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
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
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