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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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.
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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Feature
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Specifications
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Applications
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PCR, southern blotting
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Elution volume
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100–400 µl
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Format
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Spin column
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Main sample type
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Whole blood, body fluids
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Processing
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Manual (centrifugation or vacuum)
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Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein
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Genomic DNA, mitochondrial DNA, viral DNA
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Sample amount
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0.3–2 ml
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Technology
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Silica technology
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Time per run or per prep
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55 minutes
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Yield
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20–60 µg
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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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