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QIAamp DNA Stool Mini Kit

For isolation of DNA from stool

Features

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

Product Details

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.

The QIAamp® DNA Stool Mini Kit (50) (cat. no. 51504) and the InhibitEX Tablets (100) (cat. no. 19590) are no longer manufactured. Instead, we offer you the QIAamp Fast DNA Stool Mini Kit (50) (cat. no. 51604) as an alternative product.

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.

See figures

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.
See figures

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

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsPCR, blotting
Main sample typeStool samples
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinGenomic DNA, bacterial DNA, parasite DNA, viral DNA
FormatSpin column
Elution volume200 µl
TechnologySilica technology
Sample amount220 mg
Yield10–30 µg
Time per run or per prep50 minutes
ProcessingManual (centrifugation or vacuum)

Resources

Brochures & Guides (1)
Kit Handbooks (2)
For DNA purification from stool samples. Version June 2012
For DNA purification from stool samples
User-Developed Protocols (2)
The protocol describes the preservation and concentration of stool samples (in preparation for microscopic examination for intestinal parasites), using two systems from Meridian Biosciences, followed by isolation of DNA from the stool samples for pathogen detection using the QIAamp DNA Stool Mini Kit.
As starting material, 5 g soil was mixed with different amounts of Bacillus subtilis cells. Sensitivity was 5 x 103 cells/5g soil.
Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.

Publications

Assays to detect beta-tubulin codon 200 polymorphism in Trichuris trichiura and Ascaris lumbricoides.
Diawara A; Drake LJ; Suswillo RR; Kihara J; Bundy DA; Scott ME; Halpenny C; Stothard JR; Prichard RK;
PLoS Negl Trop Dis; 2009; 3 (3):e397 2009 Mar 24 PMID:19308251
Therapeutic Chlamydophila abortus and C. pecorum vaccination transiently reduces bovine mastitis associated with Chlamydophila infection.
Biesenkamp-Uhe C; Li Y; Hehnen HR; Sachse K; Kaltenboeck B;
Infect Immun; 2006; 75 (2):870-7 2006 Nov 21 PMID:17118976
Differential detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii by a single-round PCR assay.
Hamzah Z; Petmitr S; Mungthin M; Leelayoova S; Chavalitshewinkoon-Petmitr P;
J Clin Microbiol; 2006; 44 (9):3196-200 2006 Sep PMID:16954247
Molecular profiling of the Clostridium leptum subgroup in human fecal microflora by PCR-denaturing gradient gel electrophoresis and clone library analysis.
Shen J; Zhang B; Wei G; Pang X; Wei H; Li M; Zhang Y; Jia W; Zhao L;
Appl Environ Microbiol; 2006; 72 (8):5232-8 2006 Aug PMID:16885270
Isolation and characterization of a new Clostridium sp. that performs effective cellulosic waste digestion in a thermophilic methanogenic bioreactor.
Shiratori H; Ikeno H; Ayame S; Kataoka N; Miya A; Hosono K; Beppu T; Ueda K;
Appl Environ Microbiol; 2006; 72 (5):3702-9 2006 May PMID:16672520

FAQ

Are there important considerations for plasma generation and urine handling?

It is strongly advised to follow the recommendations for preparing sample material provided in the corresponding Protocol Sheet to ensure reliable results.

Plasma: It is recommended to perform plasma separation immediately after blood collection when using EDTA or citrate as anticoagulant to prevent the release of genomic DNA into the plasma fraction.

Urine: Because circulating cell-free DNA in non-stabilized urine samples is rapidly degraded after sample collection due to high nuclease activity, eluates may contain no DNA or exhibit low DNA concentration. Therefore, it is recommended to stabilize urine samples. Even when using stabilized urine, it is recommended to perform a centrifugation step immediately after stabilization to prevent the release of genomic DNA from cells. Alternatively, non-stabilized urine samples can be processed immediately after collection and centrifugation using ATL-pretreatment and automated DNA extraction as described in the corresponding Protocol Sheet.

FAQ ID - 3699
How do I safely inactivate biohazardous flow-through material?

Always dispose of potentially biohazardous solutions according to your institution’s waste-disposal guidelines. Although the lysis and binding buffers in QIAamp, DNeasy, and RNeasy kits contain chaotropic agents that can inactivate some biohazardous material, local regulations dictate the proper way to dispose of biohazards. DO NOT add bleach or acidic solutions directly to the sample-preparation waste. Guanidine hydrochloride in the sample-preparation waste can form highly reactive compounds when combined with bleach.
Please access our Material Safety Data Sheets (MSDS) online for detailed information on the reagents for each respective kit.

FAQ ID -12
How can I purify DNA from soil, food and sewage samples for PCR?

The isolation of DNA from soil, food and sewage for use in PCR can be challenging due to a high levels of inhibitors in these samples (e.g. humic acids, tannic acids, lignins, and anthocyanins) which are often co-isolated with the DNA.

The QIAamp DNA Stool Mini Kit allows isolation of genomic DNA from fecal and soil samples with the efficient removal of PCR inhibitors.

FAQ ID -118
How do I perform a DNA precipitation to concentrate my sample?
  • Add 1/10 volume of 3 M Na-Acetate pH 5.2, and 2 to 2.5 volumes of ice-cold 100% ethanol to the DNA sample
  • Mix, and store at -20°C for at least 1 hour to precipitate the DNA
  • Recover the precipitated DNA by centrifugation at full speed in a microcentrifuge for 15-20 minutes
  • Pour off the ethanol and wash the pellet twice with room-temperature 70% ethanol
  • Allow the DNA pellet to air-dry
  • resuspend the DNA in a suitable volume of sterile TE buffer or distilled water
FAQ ID -305
What can be used as an alternative to the A260 measurement for quantification of small amounts of RNA and DNA?

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728
What is the composition of buffer AE?

The composition of Buffer AE is:

  • 10 mM Tris-Cl
  • 0.5 mM EDTA; pH 9.0.
FAQ ID -730
Do I need to have EDTA in the buffer in which I am going to store my isolated genomic DNA?

EDTA chelates divalent cations which are required for nuclease activity. While the genomic DNA (gDNA) extracted using QIAGEN products, should not have any nuclease activity, it is possible to introduce nucleases during repeated long-term access of the DNA. EDTA helps to prevent any nuclease activity introduced after the genomic DNA extraction procedures.

However, if the gDNA is stored frozen at -20oC or -80oC, nuclease activity is much reduced. It may be possible to leave EDTA out of the storage buffer without negative consequences when samples are kept under these conditions, and when repeated freeze-thaw cycles are avoided.

We do recommend however that gDNA be stored in a neutral to a slightly basic buffered solution (e.g. 10 mM Tris-Cl pH 8.5 to 9.0) to prevent DNA degradation by acid hydrolysis. Note that deionized water mostly has an acidic pH.

FAQ ID -754
What is the difference between QIAGEN Protease and QIAGEN Proteinase K provided in various QIAamp Kits?

QIAGEN Proteinase K is a subtilisin-type protease, which cleaves at the carboxyl side of hydrophobic, aliphatic and aromatic amino acids. It is particularly suitable for short digestion times. It possesses a high specific activity over a wide range of temperatures and pH values with substantially increased activity at higher temperature. Soluble calcium is not essential for enzymatic activity. This means that EDTA, which is used to inhibit Mg2+-dependent enzymes such as nucleases, will not inhibit Proteinase K activity.

QIAGEN Protease is a broad-specificity Serine protease with high activity, cleaving preferentially at neutral and acidic residues. It is an economical alternative to Proteinase K for isolation of native DNA and RNA from a variety of samples.

For more information on activity of QIAGEN Protease and Proteinase K in various buffers please click here.

FAQ ID -761
Do you have a protocol for the isolation of DNA from formalin-preserved stool samples?

Yes, please follow the User-Developed Protocol 'Isolation of DNA from formalin-preserved stool samples using the QIAamp DNA Stool Mini Kit' (QA27).  Please contact Technical Service for this protocol.

FAQ ID -922
Do you have a protocol for isolation of bacterial DNA from soil?
FAQ ID -923