从血液和相关体液中纯化基因组DNA、线粒体DNA或病毒DNA,最高可达60 μg

  • 快速纯化高品质、即用型DNA
  • 无需有机提取或乙醇沉淀
  • 重复性好,产量高
  • 完全去除污染物和抑制剂

QIAamp DNA Blood Midi Kits应用硅胶模技术,用于实验室从全血、血浆、血清和其它体液中纯化DNA。QIAamp DNA Blood Midi Kit可处理0.3–2 ml新鲜或冷冻的人类全血。QIAamp Midi离心柱可在离心机或真空装置上处理(使用真空管需要额外的缓冲液AW1 [货号:19081]和AW2 [货号:19072] )。

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QIAamp DNA Blood Midi Kit (20)
For 20 DNA midipreps: 20 QIAamp Midi Spin Columns, QIAGEN Protease, Buffers, Collection Tubes (15 ml)
51183
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QIAamp DNA Blood Midi Kit (100)
For 100 DNA midipreps: 100 QIAamp Midi Spin Columns, QIAGEN Protease, Buffers, Collection Tubes (15 ml)
51185
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The QIAamp DNA Blood Midi 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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QIAamp Spin Column procedure.|Apoptotic banding in stored blood.|Long-range PCR.|Paternity testing by RFLP analysis.|

Optimized buffers lyse samples, stabilize nucleic acids, and enhance selective DNA adsorption to the QIAamp membrane. Alcohol is added and lysates loaded onto the QIAamp spin column. Wash buffers are used to remove impurities and pure, ready-to-use DNA is then eluted in water or low-salt buffer. |Genomic DNA from 8 blood samples stored at 4°C for 1 week. DNA was purified using the QIAamp DNA Blood Mini Kit. When blood is stored at 4°C, the DNA is rapidly degraded due to apoptosis; the resulting apoptotic banding pattern can clearly be seen in these samples. M1: lambda-HindIII; M2:100 bp ladder.|Amplification of a 10 kb fragment of the human ALDH1 gene from genomic DNA isolated from blood. DNA was purified using conventional methods (Phenol) or the QIAamp DNA Blood Maxi Kit (QIAamp). PCR products were sequenced directly. M: 1 kb ladder.|Three paternity cases tested with DNA purified from blood samples with the QIAamp DNA Blood Mini Kit. 1: mother; 2: child; 3: alleged father; 4: child + alleged father. M: markers. Each lane had 3 µg of DNA loaded. Outcome: A: alleged father excluded; B & C: alleged father confirmed.
(Data kindly provided by J. James, Gene Proof Technologies, Nashville, TN, USA.)|

Performance

QIAamp DNA Blood Kits根据样本存储条件和新鲜程度不同,可纯化获得200 bp–50 kb的DNA(参见"Apoptotic banding in stored blood")。纯化的DNA适用于长片段PCR扩增(参见"Long-range PCR")和RFLP分析等等,如亲子鉴定(参见"Paternity testing by RFLP analysis")。

使用QIAamp DNA Blood Midi Kit获得DNA的回收率可高达94.5%,依初始细胞密度而定(见表)。

从不同细胞密度的人类全血中纯化的DNA产量
白细胞/ ml DNA产量(µg) DNA回收率(%)
2.5 x 105 3.0 92.3
1.0 x 106 11.0 91.7
5.0 x 106 62.4 94.5
1.0 x 107 116.3 88.1
从2 ml人类全血中纯化得到基因组DNA,并用300 µl洗脱液洗脱。第一次的洗出液再次上样到离心柱并离心(即:再洗脱)。假设一个白细胞包含6.6 pgDNA,计算DNA回收率。 
Principle

无需苯酚-氯仿抽提。核酸特异性结合到QIAamp硅胶膜上,污染物流走。PCR抑制剂,如:二价阳离子和蛋白,可通过两步有效的洗涤被完全去除,结合在离心柱上的纯核酸可用水或试剂盒中的缓冲液洗脱。QIAamp DNA Blood技术从全血或相关体液纯化得到的基因组、线粒体或病毒DNA,可即用于PCR和印迹实验中。

Procedure

QIAamp DNA Blood Midi Kit采用快速离心柱操作简化了从全血或相关体液中分离DNA的流程(参见"procedure")。该试剂盒可处理带有EDTA、柠檬酸盐和肝素等常规抗凝剂的新鲜或冷冻的全血样本。QIAamp DNA Blood Midi Kit可处理0.3–2 ml的样本,制备时间55分钟。可从1 ml健康全血中获得20–60 µg DNA,洗脱体积100–400 µl。

QIAamp DNA Blood Kits提供全面的操作手册,包括从不同来源的样本中纯化DNA的操作流程。QIAamp样本制备技术已完全经过验证。

Applications

QIAamp DNA Blood Midi Kit使用经验证的QIAamp技术,采用带有15 ml收集管的Midi离心柱从不同来源的样本中纯化DNA。样本来源包括:

新鲜和冷冻的全血或白膜层
血浆或血清
骨髓
淋巴细胞
体液
Feature
Specifications
Applications PCR, southern blotting
Elution volume 100–400 µl
Format Spin column
Main sample type Whole blood, body fluids
Processing Manual (centrifugation or vacuum)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein Genomic DNA, mitochondrial DNA, viral DNA
Sample amount 0.3–2 ml
Technology Silica technology
Time per run or per prep 55 minutes
Yield 20–60 µg

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Kit Handbooks
1
For large-scale genomic and viral DNA purification from whole blood, plasma, serum, body fluids, lymphocytes
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User-Developed Protocols
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As starting material, 5 g soil was mixed with different amounts of Bacillus subtilis cells. Sensitivity was 5 x 103 cells/5g soil.
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References
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