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With the development of molecular biology, stool, a new non-invasive sample, has been widely used in the research of animal molecular genetics, population ecology, behavioral ecology and some intestinal disease diagnosis. Stool samples includes gut microbial DNA, food residue sample DNA, and alimentary tract exfoliated cell DNA.
The primary problem encountered when using stool sample for molecular biology research is the low content of exfoliated cells in the digestive tract and a certain degree of degradation of genetic material in stool. Another issue in molecular scatology research based on PCR is the presence of a large number of inhibitors in stool that can affect Taq enzyme activity, leading to downstream detection inactivation. These inhibitors include polysaccharides, plant polysaccharides, bile acids, bile salts, bile pigments, digestive juices, mucus, etc. Therefore, selecting appropriate extraction methods to obtain high-quality DNA is the key to successful downstream detection of stool DNA.
At present, the pretreatment methods used in the laboratory, such as phenol/chloroform extraction, cetyltrimethyl bromide (CTAB) lysis, and guanidine isothiocyanate lysis, lack universality in different species, and the success rate of extracting DNA for PCR amplification is also very low. The HiPure Stool DNA Kit provided by Magen Company has opened up a new approach for DNA extraction from stool samples with good universality, high cost-effectiveness, high yield and purification. The reagent kit adopts a unique solution system and inhibitory factor adsorbent, which can efficiently remove various impurities in stool samples. The purified DNA can be directly used for PCR, quantitative PCR and other applications.
This product allows rapid and reliable isolation of high-quality genomic DNA from various stool samples. Up to 100 mg soil samples can be processed in 60 minute. The system combines the reversible nucleic acid binding properties of HiPure matrix with the speed and versatility of spin column technology to eliminate PCR inhibiting compounds such as humic acid from soil samples. Purified DNA is suitable for PCR, restriction digestion, and next-generation sequencing. There are no organic extractions thus reducing plastic waste and hands-on time to allow multiple samples to be processed in parallel.
Details
Specifications
Features
Specifications
Main Functions
Isolation total DNA from 50-100mg stool samples
Applications
PCR, Southern Blot, enzyme digestion and NGS, etc.
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Stool
Sample amount
50-100mg
Yield
3-15μg
Elution volume
≥30μl
Time per run
≤60 minutes
Liquid carrying volume per column
750μl
Binding yield of column
100μg
Principle
Stool sample is homogenized and then treated in a specially formulated buffer containing detergent to lyse bacteria, yeast, and fungal samples. Humic acid, proteins, polysaccharides, and other contaminants are removed using our proprietary Absorber Solution. Binding conditions are then adjusted and the sample is applied to a DNA Mini Column. Two rapid wash steps remove trace contaminants and pure DNA is eluted in low ionic strength buffer. Purified DNA can be directly used in downstream applications without the need for further purification.
Advantages
High purity – unique adsorbent can completely remove inhibitory factors
High concentration – maximum extraction of total DNA from stool samples
High recovery – DNA can be recovered at the level of PG
Good repeatability – silica technology can obtain ideal results every time
Kit Contents
Contents
IVD3141
Purification Times
50 Preps
HiPure DNA Mini Columns II
50
2ml Collection Tubes
50
2ml Bead Tubes
50
Proteinase K
24 mg
Protease Dissolve Buffer
1.8 ml
Buffer SPL
40 ml
Buffer PCI
40 ml
Buffer AL
20 ml
Buffer GW1
22 ml
Buffer GW2
20 ml
Buffer AE
15 ml
Storage and Stability
Proteinase K and Buffer PCI should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2–8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.
Document
With the development of molecular biology, stool, a new non-invasive sample, has been widely used in the research of animal molecular genetics, population ecology, behavioral ecology and some intestinal disease diagnosis. Stool samples includes gut microbial DNA, food residue sample DNA, and alimentary tract exfoliated cell DNA.
Storage term: stored at ≤-20℃,keep away from light, avoid heavy weight and repeated freezing and thawing.
Term of Validity: 14 months
Isothermal Nucleic Acid Principle Summary
The kit is based on rapid nucleic acid amplification technology at room temperature and constant temperature, its principle is that at room and constant temperature, the recombinase and primer form a protein/single-stranded nucleotide complex Rec/ssDNA, and invade the double-stranded DNA template with the help of auxiliary proteins and single-stranded binding protein SSB; then form a D-loop region at the invasion point and start to scan the DNA duplex, after finding the target region complementary to the primer and disintegration of the complex Rec/ssDNA, the polymerase also binds to the 3′ end of the primer to start the chain extension.
Isothermal Nucleic Acid Product Features
1/ High sensitivity and specificity, short reaction time.
2/ The reagent form is freeze-dried, stable and easy to operate.
3/ The reaction can be operated by metal bath and water bath pot without purchasing expensive PCR apparatus.
Technical Parameters:
Parameters
Details
Product Name
DNA Isothermal Amplification Kit Basic
Manufacturer
Amp-future
Storage Temperature
-20°C
Kit Components
Enzymes, Buffers ,Reagents
Packaging
48 Tests/box
Detection Limit
500-1000copies/µL
Shipping
ICE
Test Time
5-20mins
Isothermal Nucleic Acid Applications
Suitable for DNA isothermal rapid amplification kit(Basic type)
Primer: Require pair of nucleotide primers with the length of 25-35 bp.
Colloidal gold probe:Require a sequence of 46-52nt in length
DNA basic kit reaction temperature is 39 to 42℃ and time is 5-20 minutes.
Notes
1/ Please avoid nucleic acid contamination and set blank control during reaction due to the high sensitivity of the kit.
2/ Please take out the required quantity of MIRA reaction units for the experiment, and put the rest under storage conditions when performing the experiment.