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)

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
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