For isolation of genomic, mitochondrial, bacterial, parasite or viral DNA
- Rapid purification of high-quality, ready-to-use DNA
- Consistent, high yields
- Complete removal of contaminants and inhibitors
The QIAamp DNA Mini Kit provides silica-membrane-based nucleic acid purification from tissues, swabs, CSF, blood, body fluids, or washed cells from urine. No mechanical homogenization is necessary as the tissues are lysed enzymatically. The convenient spin-column procedure reduces hands-on preparation time to 20 minutes. Purification of DNA using the QIAamp DNA Mini Kit can be automated on the QIAcube Connect.
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QIAamp DNA Mini Kit (50)
For 50 DNA preps: 50 QIAamp Mini Spin Columns, QIAGEN Proteinase K, Reagents, Buffers, Collection Tubes (2 ml)
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51304
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QIAamp DNA Mini Kit (250)
For 250 DNA preps: 250 QIAamp Mini Spin Columns, QIAGEN Proteinase K, Reagents, Buffers, Collection Tubes (2 ml)
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51306
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The QIAamp DNA Mini Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Purification of up to 50 kb genomic DNA.|The QIAcube.|
Size distribution of DNA prepared with QIAamp Kits from the indicated sources (3 µg per lane).||
Performance
DNA purified using the QIAamp DNA Mini Kit can be used in a wide range of downstream applications, including PCR and quantitative real-time PCR, Southern blotting, SNP and STR genotyping, and pharmocogenomic research.
The QIAamp DNA Mini Kit yields DNA sized up to 50 kb (see figure "Purification of up to 50 kb genomic DNA"). DNA of this length denatures completely and has the highest amplification efficiency. Yields of nucleic acids or DNA depend on the starting material (see table).
QIAamp sample preparation technology is fully licensed, allowing QIAamp purified nucleic acids to be used in any molecular assay or other downstream application without risk of patent infringement.
| Blood |
200 µl |
4–12 |
4–12 |
| Buffy coat |
200 µl |
25–50 |
25–50 |
| Cells |
106 |
20–30 |
15–20 |
| Liver |
25 mg |
60–115 |
10–30 |
| Brain |
25 mg |
35–60 |
15–30 |
| Lung |
25 mg |
25–45 |
5–10 |
| Heart |
25 mg |
15–40 |
5–10 |
| Kidney |
25 mg |
40–85 |
15–30 |
| Spleen |
10 mg |
25–45 |
5–30 |
Principle
The QIAamp DNA Mini Kit simplifies isolation of DNA from human tissue samples with fast spin-column or vacuum procedures. 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 DNA to be eluted in either water or a buffer provided with the kit. QIAamp DNA technology yields genomic, mitochondrial, bacterial, parasite, or viral DNA from human tissue samples ready to use in PCR and blotting procedures.
Procedure
Optimized buffers and enzymes 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.
No mechanical homogenization is necessary as the tissues are lysed enzymatically, and the convenient spin-column procedure means that hands-on preparation time is only 20 minutes (lysis times differ according to the sample source). Samples can be processed using either a microcentrifuge or, if blood or other body fluids are being processed, using the QIAvac 24 Plus or QIAvac 6S vacuum manifold. In addition, the rigorous lysis procedure employed makes the QIAamp DNA Mini Kit ideal for purification of genomic DNA from bacteria or parasites. To further reduce hands-on time, genomic DNA purification may be automated on the QIAcube.
Vacuum processing
Blood or other body fluids can be processed by vacuum, instead of centrifugation, for greater speed and convenience in DNA purification. QIAamp Mini spin columns are accommodated on the QIAvac 24 Plus manifold using VacValves and VacConnectors. VacValves should be used if sample flow rates differ significantly, to ensure consistent vacuum. Disposable VacConnectors are used to avoid any cross-contamination. Use of VacConnectors also allows these QIAamp spin procedures to be performed on QIAvac 6S with QIAvac Luer Adapters.
Applications
The QIAamp DNA Mini Kit is ideal for purification of DNA from most commonly used human tissue samples, including muscle, liver, heart, brain, bone marrow, and other tissues, swabs (buccal, eye, nasal, pharyngeal, and others), CSF, blood, body fluids, and washed cells from urine. DNA can be purified from up to 25 mg tissue or from up to 200 µl fluid in 20 minutes, and eluted in 50–200 µl.
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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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50–200 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, tissue, cells
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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, bacterial DNA, parasite DNA, viral DNA
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Sample amount
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200 µl/25 mg/5 x 10e6
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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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20 minutes
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Yield
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4–30 µg
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For parallel preparation of genomic, bacterial, or viral DNA or viral RNA from more than 24 samples, we recommend using QIAamp® Spin Columns with 4-6 ml collection tubes. Any tube with an inner diameter of 9-10 mm (outer diameter 11-12 mm) is suitable.
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The protocol can be used for fresh or frozen semen samples with equal efficiency. Frozen samples must be thawed thoroughly before use. Please note that lysis time will vary depending on the size and density of the source material.
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These procedures have been used successfully for isolation of genomic DNA from Aspergillus and Candida species, from both fungal cultures and blood.
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This protocol has only been tested with ‘soft’ tissues (e.g., liver, spleen, thymus, heart, kidney, and brain) and may not work with ‘hard’ tissues (e.g., bone, teeth, and skin).
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