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Product description

 

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.
Principle

 

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 from Various Animal Tissues", see table "Typical DNA Yields from Animal Tissues Using DNeasy Blood & Tissue Kits").

Typical DNA Yields from Animal Tissues Using DNeasy Blood & Tissue Kits

High-Quality DNA from Various Animal Tissues

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.

 

The efficient DNeasy Blood & Tissue procedure enables high yields of total DNA from animal blood and tissue samples (table "Typical DNA Yields from Animal Tissues Using DNeasy Blood & Tissue Kits", see figure "Typical DNA Yields from Animal Blood Using DNeasy Blood & Tissue Kits"). Optimized protocols ensure high yields from nonstandard samples, such as animal hair (see figure "Genotyping with DNA from Horse Hairs"), 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 DNA Yields from Animal Blood Using DNeasy Blood & Tissue Kits

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.

 

Genotyping with DNA from Horse Hairs

 

DNA was purified from horse 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.

 

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 spin columns are prepackaged in collection tubes and individually sealed, providing convenience and safety. The DNeasy 96 Blood & Tissue Kit provides high-throughput processing in a 96-well format using the QIAGEN 96-Well-Plate Centrifugation System. For standardized automated purification, comparable results are obtained using the BioSprint 15 DNA Blood Kit and the BioSprint 96 DNA Blood Kit on BioSprint workstations (please inquire).

 

 

Applications

 

DNeasy Blood & Tissue Kits provide reliable results — from laboratory analysis of transgenic mice (see figure "High-Throughput DNA Purification Using the DNeasy 96 Blood & Tissue Kit") to livestock breeding programs (see figure "Genotyping of Pigs for Quality Management of Livestock") and pedigree genotyping. Routine testing applications can be easily scaled up using the DNeasy 96 Blood & Tissue Kit (see figure "High-Throughput DNA Purification Using the DNeasy 96 Blood & Tissue Kit").

High-Throughput DNA Purification Using the DNeasy 96 Blood & Tissue Kit

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.

 

Genotyping of Pigs for Quality Management of Livestock

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). (Data kindly provided by Dr Werner Brabetz, Biotype AG, Dresden, Germany; info@biotype.de; www.biotype.de )

 

Purified DNA is free from PCR inhibitors, enabling sensitive detection in standard, multiplex (see figure "Efficient 16plex PCR over a Range of Template Amounts"), and real-time PCR (see figure "Real-Time PCR Detection of Borrelia DNA in Ticks"). 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

Efficient 16plex PCR over a Range of Template Amounts

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.

 

Real-Time PCR Detection of Borrelia DNA in Ticks

 

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.