For isolation of DNA from stool
  • Rapid isolation of high-quality, ready-to-use DNA
  • No organic extraction or alcohol precipitation
  • Consistent, high yields
  • Complete removal of contaminants and inhibitors

The QIAamp DNA Stool Mini Kit provides silica membrane-based purification of up to 30 μg genomic, bacterial, viral, and parasite DNA from fresh or frozen human stool or other sample types with high concentrations of PCR inhibitors. The combined action of InhibitEX, a unique adsorption resin, and an optimized buffer leads to removal of PCR inhibitors. The convenient QIAamp spin-column procedure provides purification in only 50 minutes. Purification of DNA using the QIAamp DNA Stool Mini Kit can be automated on the QIAcube.

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The QIAamp DNA Stool Mini 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 DNA Stool Mini procedure.|Removal of PCR inhibitors.|
|DNA was purified from 19 stool samples using [A] a conventional silica-based purification technique and [B] the QIAamp DNA Stool Mini Kit. To show whether inhibitors were present in the purified eluates, 5 µl of each eluate was added to PCR mixes with a template of 10 pg plasmid containing a GFP gene. Amplification of the GFP gene was successful in the presence of all QIAamp eluates whereas only 2 amplification reactions were successful in the presence of eluates prepared using the conventional technique containing the green fluorescent protein. M: markers; C: positive PCR control.|
Performance

The QIAamp DNA Stool Mini Kit allows rapid purification of DNA from fresh or frozen human stool or other sample types with high concentrations of PCR inhibitors. Up to 220 mg stool can be processed routinely, and larger amounts can be processed with additional Buffer ASL. Typical yields of 10–30 µg are obtained in 50 minutes, and DNA is eluted in 200 µl.  The purified DNA is sized up to 50 kb. DNA of this length denatures completely and has the highest amplification efficiency. Highly pure DNA is ready for direct use in downstream amplification reactions (see figure "Removal of PCR inhibitors").

DNA that has been purified using the QIAamp DNA Stool Mini Kit can be used in a wide range of downstream applications, including PCR and quantitative real-time PCR, infectious disease research, and screening.

Principle

DNA binds specifically to the QIAamp silica-gel membrane while contaminants pass through. No phenol–chloroform extraction is required. PCR inhibitors are removed by the combined action of InhibitEX, a unique adsorption resin, and an optimized buffer. Rigorous lysis using proteinase K ensures high yields of all types of DNA common in stool, including colorectal epithelial cells, bacteria, viruses and other gastrointestinal pathogens.

Highly pure DNA ready for direct use in downstream amplification reactions is purified in about 50 minutes. QIAamp sample preparation technology is fully licensed.

Procedure
The QIAamp DNA Stool Mini Kit simplifies isolation of DNA from stool with a fast spin-column procedure (see flowchart "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. PCR inhibitors are removed by the combined action of InhibitEX and an optimized buffer. Proteinase K lysis ensures high yields of all types of DNA common in stool. Remaining impurities are efficiently removed in two wash steps. Amplification-ready DNA is then eluted in low-salt buffer.
Applications

The QIAamp DNA Stool Mini Kit is for isolation of genomic, bacterial, viral, and parasite DNA from the following sample types:

Fresh stool
Frozen stool
Other sample types with high concentrations of PCR inhibitors
Feature
Specifications
Applications PCR, blotting
Elution volume 200 µl
Format Spin column
Main sample type Stool samples
Processing Manual (centrifugation or vacuum)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein Genomic DNA, bacterial DNA, parasite DNA, viral DNA
Sample amount 220 mg
Technology Silica technology
Time per run or per prep 50 minutes
Yield 10–30 µg