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 theQIAcube Connect.
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
DNA yields.
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.
DNeasy Mini and 96 procedures.
Efficient 16plex PCR.
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.
High-quality DNA.
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.
Genotyping of horses.
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.
High-throughput purification.
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.
Real-time PCR.
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.
Genotyping of pigs.
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).
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
Source
Amount
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").
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.
Applications
DNeasy Blood & Tissue Kits provide high-quality DNA, ready to use in all downstream assays, including applications in:
For 12 x 96 total and cytoplasmic RNA preps: 12 RNeasy 96 Plates, Elution Microtubes CL, Caps, S-Blocks, AirPore Tape Sheets, RNase-Free Reagents and Buffers
For 4 x 96 total and cytoplasmic RNA preps: 4 RNeasy 96 Plates, Elution Microtubes CL, Caps, S-Blocks, AirPore Tape Sheets, RNase-Free Reagents and Buffers
2.5 ml (100 mg/ml; 7000 units/ml, solution)
Unit definition: That amount of enzyme causing the hydrolysis of RNA at a rate such that k (velocity constant) equals unity (Kunitz units) at 25°C and pH 5.0.
