DNeasy Blood & Tissue Kits

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

  • 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 Kits provide fast and easy silica-based DNA purification 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 purification of high-quality DNA for life science, genotyping, and veterinary pathogen research applications. Purification of DNA using the DNeasy Blood & Tissue Kit can be automated on the QIAcube Connect.

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製品名 Cat. no. List price:
DNeasy 96 Blood and Tissue Kit (4)
For 4 x 96 DNA minipreps: 4 DNeasy 96 Plates, Proteinase K, Buffers, S-Blocks, AirPore Tape Sheets, Collection Microtubes (1.2 ml), Elution Microtubes RS, Caps, 96-Well Plate Registers
69581
¥156,000
DNeasy 96 Blood and Tissue Kit (12)
For 12 x 96 DNA minipreps: 12 DNeasy 96 Plates, Proteinase K, Buffers, S-Blocks, AirPore Tape Sheets, Collection Microtubes (1.2 ml), Elution Microtubes RS, Caps, 96-Well Plate Registers
69582
¥420,000
DNeasy Blood and Tissue Kit (50)
50 DNeasy Mini Spin Columns, Proteinase K, Buffers, Collection Tubes (2 ml)
69504
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DNeasy Blood and Tissue Kit (250)
250 DNeasy Mini Spin Columns, Proteinase K, Buffers, Collection Tubes (2 ml)
69506
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DNeasy Blood & Tissue Kits  は分子生物学実験用です。疾病の診断、治療または予防の目的には使用することはできません。


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DNA yields.|DNeasy Mini and 96 procedures.|Efficient 16plex PCR.|High-quality DNA.|Genotyping of horses.|High-throughput purification.|Real-time PCR.|Genotyping of pigs.|
DNA was purified from 100 µl whole blood using the DNeasy Blood & Tissue Kit. Goat: DNA was purified from 50 µl goat whole blood. Using more than 50 µl goat blood gave no significant increase in DNA yield. Chicken: DNA was purified from 5 µl chicken whole blood. Bird blood contains nucleated erythrocytes, giving higher DNA yields than mammalian blood.||Genomic DNA was purified from human K562 cells using the DNeasy Blood & Tissue Kit. Multiplex PCRs were performed in duplicate using the QIAGEN Multiplex PCR Kit with the indicated amount of DNA and 16 pairs of primers. M: 100 bp ladder.|DNA was purified from the indicated animal tissues using the DNeasy Blood & Tissue Kit and analyzed by agarose gel electrophoresis. For each sample, 0.5 µg DNA was analyzed undigested (left) or 1 µg DNA was digested with EcoRI (right). M: lambda-HindIII.|DNA was purified from hair of 3 brown horses and analyzed for coat-color genotypes using the MSH and ASIP PCR systems. [A] MSH PCR products were digested with TaqI. Horses 1 and 3 were identified as Ee heterozygotes, carrying the recessive e allele for red coat color. [B] Analysis of ASIP PCR products identify horses 1 and 2 as Aa heterozygotes, carrying the recessive a allele for black coat color. +: Positive Ee aa control DNA from a black horse. -: Negative PCR control. M: markers.|DNA was purified from 24 mouse tails and from 24 rat tails using the DNeasy 96 Blood & Tissue Kit. DNA was analyzed on a 1% agarose gel. M: lambda-HindIII.|Borrelia sp. is a pathogen carried by ticks that causes Lyme disease in humans and other mammals. DNA was purified from ticks using the DNeasy Blood & Tissue Kit. Borrelia DNA was detected in real-time PCR using the artus Borrelia LC PCR Kit on the LightCycler system. The lowest positive curve corresponds to 5 copies per PCR.|DNA was purified from pig ear punches using the DNeasy Blood & Tissue Kit and amplified using the Animaltype Pig PCR Amplification Kit from Biotype AG and QIAGEN HotStarTaq DNA Polymerase. The electropherogram shows the genetic fingerprint for 12 polymorphic tetranucleotide microsatellites and one marker for gender determination (SBH23).
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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").

Principe

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.

Procédure

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.

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
Feature
Specifications
Applications PCR, real-time PCR, genotyping
Elution volume 100–200 µl
Format 96-well plate, spin column
Main sample type Blood, tissue
Processing Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein DNA
Sample amount 100 µl/25 mg
Technology Silica technology
Time per run or per prep 20 minutes – 1 hour
Yield 6 µg/30 µg

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Supplementary Protocols
4
Kit Handbooks
1
For purification of total DNA from animal blood, animal tissue, rodent tails, ear punches, cultured cells, fixed tissue, bacteria, insects

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Typical DNA Yields from Animal Blood Using DNeasy Blood & Tissue Kits
DNA収量
DNeasy Blood & Tissue Kitを用いて100 µlの全血からDNAを精製した。ヤギ:ヤギの全血50 µlからDNAを精製した。ヤギの血液50 µl以上を用いてもDNA収量に顕著な増加はなかった。ニワトリ:ニワトリの全血5 µlからDNAを精製した。鳥類の血液は有核赤血球を含んでいるため、哺乳動物の血液から精製されるDNAより収量が多い。
DNeasy Tissue and DNeasy 96 Tissue Procedure
DNeasy Miniおよび96精製法
Efficient 16plex PCR over a range of template amounts
効率的な16plex PCR
DNeasy Blood & Tissue Kit を用いてヒトK562細胞からゲノムDNAを精製した。表記の量のDNAと16種類のプライマーペア、QIAGEN Multiplex PCR Kitを用いてduplicateでマルチプレックスPCRを行なった。M:100 bpラダー。
High-Quality DNA from Various Animal Tissues
高品質なDNA
表記の動物組織からDNeasy Blood & Tissue Kitを用いてDNAを精製し、アガロースゲル電気泳動により解析した。各サンプルの左レーンには未処理のDNA 0.5 µg、右のレーンにはEcoRIで切断したDNA 1 µgをロードして、解析した。M: lambda-HindIII。
Genotyping with DNA from Horse Hairs
ウマのジェノタイピング
栗毛のウマ3頭の毛からDNAを精製し、MSHおよびASIP PCRを用いて毛色のジェノタイプを解析した。[A] MSH PCR産物をTaqIで切断した。ウマ1、3は赤毛の劣性e対立遺伝子を持つヘテロ型のEeと同定された。[B] ASIP PCR産物を解析した結果、ウマ1、2は黒毛の劣性a対立遺伝子を持つヘテロ型のAaと同定された。+:黒毛のウマから精製したポジティブなEe aaコントロールDNA。-:ネガティブPCRコントロール。M:マーカー。
High-Throughput DNA Purification Using the DNeasy 96 Blood & Tissue Kit
ハイスループット精製
DNeasy 96 Blood & Tissue Kitを用いてマウスの尾およびラットの尾、それぞれ24匹分からDNAを精製した。DNAを1%アガロースゲル電気泳動で解析した。M: lambda-HindIII。
Variable elution volumes allow a range of yields and concentrations
リアルタイムPCR
Borrelia sp.はダニが媒介する病原体でヒトやその他の哺乳動物にライム病を引き起こす。DNeasy Blood & Tissue Kitを用いてダニからDNAを精製した。artus Borrelia LC PCR KitとLightCyclerシステムを用いてBorrelia DNAをリアルタイムPCRで検出した。最低量のポジティブ曲線はPCRあたり5コピーに相当する。
Genotyping of Pigs for Quality Management of Livestock
ブタのジェノタイピング
DNeasy Blood & Tissue Kitを用いてブタ耳穿孔からDNAを精製し、Biotype AG社のAnimaltype Pig PCR Amplification KitおよびQIAGEN HotStarTaq DNA Polymeraseを用いて増幅した。エレクトロフェログラムは12種類の多型テトラヌクレオチドマイクロサテライトと1種類の性別決定用マーカー(SBH23)の遺伝子フィンガープリントを示している。