Introduction

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

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

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.

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.

Description 

The FluoroStain™ Protein Fluorescent Staining Dye (Red, 1000×) is designed to substitute the common Coomassie Blue protein staining method, offering greater sensitivity and ease of operation. Unlike Coomassie Blue stain, the FluoroStain Protein Fluorescent Staining Dye binds to protein with high specificity, making destaining process an option rather than a requirement. With further reduction of background signals via destaining process, the FluoroStain™ is capable of achieving detection level parallel to silver staining without specialized imaging equipment, making it one of the most sensitive dyes available. In addition to its remarkable sensitivity, the FluoroStain™ Protein Fluorescent Staining Dye (Red, 1000×) brings a more reliable and safer user experience, since the stained gel can be visualized with blue-light illumination, avoiding the risk of skin/ eye damage caused by UV light. For best result, we suggest using B-BOX™ Blue Light LED epi-illuminator to visualize and analyze the gel stained with FluoroStain Protein Fluorescent Staining Dye (Red, 1000×). The FluoroStain™ Protein Fluorescent Staining Dye is compatible to the analysis of mass spectra, i.e. LC-MS/MS, MALDI-TOF, etc.

Spectral Characteristics

When it is bound with bovine serum albumin (BSA), the fluorescent emission of FluoroStain Protein Fluorescent Staining Dye can be excited by UV and blue light sources, with excitation peaks around 369 and 517 nm and emission at 605 nm. In absence of BSA, FluoroStain Protein Fluorescent Staining Dye shows ignorable fluorescence as compared with protein-bound form, therefore giving a clear background for photographic analysis.

These spectral characteristics made this fluorescent dye compatible with a wide variety of gel reading facilities, including UV/ blue light epi- and transilluminator, argon laser and mercury-arc lamp excitation gel scanners.

Storage 

Protected from light 
-20°C for 24 months 

The FluoroStain™ Protein Fluorescent Staining Dye (Red, 1000×) is designed to substitute the common Coomassie Blue protein staining method, offering greater sensitivity and ease of operation. Unlike Coomassie Blue stain, the FluoroStain Protein Fluorescent Staining Dye binds to protein with high specificity, making destaining process an option rather than a requirement. With further reduction of background signals via destaining process, the FluoroStain™ is capable of achieving detection level parallel to silver staining without specialized imaging equipment, making it one of the most sensitive dyes available. In addition to its remarkable sensitivity, the FluoroStain™ Protein Fluorescent Staining Dye (Red, 1000×) brings a more reliable and safer user experience, since the stained gel can be visualized with blue-light illumination, avoiding the risk of skin/ eye damage caused by UV light. For best result, we suggest using B-BOX™ Blue Light LED epi-illuminator to visualize and analyze the gel stained with FluoroStain Protein Fluorescent Staining Dye (Red, 1000×). The FluoroStain™ Protein Fluorescent Staining Dye is compatible to the analysis of mass spectra, i.e. LC-MS/MS, MALDI-TOF, etc.