从动物血液和组织、细胞、酵母、细菌或者病毒中纯化总DNA

  • 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

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。

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DNeasy 96 Blood & 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
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DNeasy 96 Blood & 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
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DNeasy Blood & Tissue Kit (50)
50 DNeasy Mini Spin Columns, Proteinase K, Buffers, Collection Tubes (2 ml)
69504
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DNeasy Blood & Tissue Kit (250)
250 DNeasy Mini Spin Columns, Proteinase K, Buffers, Collection Tubes (2 ml)
69506
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The DNeasy Blood & Tissue Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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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

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。

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。

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
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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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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Supplementary Protocols
4
References
0