QIAamp DNA Blood Midi Kit
For purification of up to 60 µg genomic, mitochondrial, or viral DNA from blood and related body fluids
- Rapid purification of high-quality, ready-to-use DNA
- No organic extraction or alcohol precipitation
- Consistent, high yields
- Complete removal of contaminants and inhibitors
QIAamp DNA Blood Midi Kits provide silica-membrane-based DNA purification for labs that process whole blood, plasma, serum, and other body fluids. The QIAamp DNA Blood Midi Kit is designed for processing 0.3–2 ml fresh or frozen human whole blood. The QIAamp Midi spin columns can be easily processed in a centrifuge or on vacuum manifolds (additional Buffer AW1 [Cat. no. 19081] and Buffer AW2 [Cat. no. 19072] are required for use with vacuum manifolds).
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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)
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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)
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51185
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QIAamp DNA Blood Midi Kit适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。
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 yield DNA sized from 200 bp to 50 kb, depending on the age and storage of samples (see figure "Apoptotic banding in stored blood"). The purified DNA is suitable for long-range PCR amplification (see figure "Long-range PCR") and restriction fragment length polymorphism (RFLP) analysis used, for example, for paternity testing (see figure "Paternity testing by RFLP analysis").
The QIAamp DNA Blood Midi Kit yields up to 94.5% recovery DNA, depending on the starting cell densities (see table).
| 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 |
Principle
No phenol–chloroform extraction is required. DNA binds specifically to the QIAamp silica-gel membrane while contaminants pass through. PCR inhibitors, such as divalent cations and proteins, are completely removed in two efficient wash steps, leaving pure nucleic acid to be eluted in either water or a buffer provided with the kit. QIAamp DNA Blood technology yields genomic, mitochondrial, or viral DNA from blood and related body fluids ready to use in PCR and blotting procedures.
Procedure
The QIAamp DNA Blood Midi Kit simplifies isolation of DNA from blood and related body fluids with a fast spin-column procedure (see flowchart "procedure"). Fresh and frozen whole blood with common anticoagulants, such as EDTA, citrate, or heparin, may be processed. The QIAamp DNA Blood Midi Kit processes sample sizes of 0.3–2 ml, with a preparation time of 55 minutes. The typical yield from 1 ml healthy whole blood is 20–60 µg, with an elution volume of 100–400 µl.
QIAamp DNA Blood Kits are supplied with a comprehensive handbook containing protocols for DNA isolation from a number of sample sources. QIAamp sample preparation technology is fully licensed.
Applications
The QIAamp DNA Blood Midi Kit provides proven QIAamp technology for purification of DNA from a variatey of materials using Midi spin columns with 15 ml collection tubes. Sample sources include:
Fresh and frozen whole blood or buffy coat
Plasma or serum
Bone marrow
Lymphocytes
Body fluids
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特点
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参数
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应用
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PCR, southern blotting
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洗脱体积
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100–400 µl
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规格
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Spin column
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主要样本类型
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Whole blood, body fluids
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处理
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Manual (centrifugation or vacuum)
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纯化总RNA、miRNA、poly A+ mRNA、DNA或蛋白
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Genomic DNA, mitochondrial DNA, viral DNA
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样本量
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0.3–2 ml
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技术
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Silica technology
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每次运行或制备时间
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55 minutes
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产量
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20–60 µg
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Second edition — innovative tools
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For large-scale genomic and viral DNA purification from whole blood, plasma, serum, body fluids, lymphocytes
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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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图片
QIAamp Spin Column实验流程。
采用优化的缓冲液裂解样本、稳定核酸并增强DNA在QIAamp膜上的选择性吸附。加入乙醇,并将裂解物上样于QIAamp离心柱。使用洗涤缓冲液去除杂质,然后用水或低盐缓冲液洗脱纯的即用型DNA。
储存的血液中的细胞凋亡带型。
将8个血液样本的基因组DNA置于4°C保存1周。使用QIAamp DNA Blood Mini Kit纯化DNA。当血液置于4°C保存时,由于细胞凋亡作用,DNA会迅速降解;在这些样本中可以清晰地观察到形成的凋亡带型。M1:lambda-HindIII;M2:100 bp分子量标准。
长片段PCR。
从血液中分离的基因组DNA中的人ALDH1基因的10 kb片段的扩增。使用传统方法(酚)或QIAamp DNA Blood Maxi Kit(QIAamp)纯化DNA。直接测序PCR产物。M:1 kb分子量标准。
采用RFLP分析进行亲子鉴定。
在三个亲子鉴定案例中,检测使用QIAamp DNA Blood Mini Kit从血液样本中纯化的DNA。1: 母亲;2:孩子;3:疑父;4:孩子+疑父。M:分子量标准。每个泳道上样3 µg DNA。结果:A:被排除的疑父;B & C:确认的疑父。(数据由美国田纳西州纳什维尔Gene Proof Technologies的J. James提供。)
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