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

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	D314102	D314103
Purification Times	50 Preps	250 Preps
HiPure DNA Mini Columns II	50	250
2ml Collection Tubes	50	250
2ml Bead Tubes	50	250
Proteinase K	24 mg	120 mg
Protease Dissolve Buffer	1.8 ml	10 ml
Buffer SPL	40 ml	200 ml
Buffer PCI	40 ml	200 ml
Buffer AL	20 ml	80 ml
Buffer GW1	22 ml	88 ml
Buffer GW2	20 ml	2 x 50  ml
Buffer AE	15 ml	30 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.

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.

Overview

• Milk samples are stable for 1 month at room temperature (or 1 week at 37°C) in the Preservation Solution
• Fast and easy processing using a rapid spin-column format
• DNA can be isolated and detected from as little as 100 µL of milk
• Isolate high quality genomic DNA

Norgen’s Milk DNA Preservation and Isolation Kit is an all-in-one solution designed for 1) preservation of DNA in milk samples at ambient temperature; and 2) isolation of high quality DNA within a laboratory setting. The kit is provided with a Preservation Solution that allows for either immediate processing of milk samples, or allows for the storage of the milk samples for up to 1 month at room temperature prior to DNA isolation. Genomic DNA extracted from somatic cells and bacteria found in milk can be used in various applications. The isolated DNA can be used for the detection of biomarkers to diagnose a disease, follow the diseases progress or monitor the effects of a particular treatment. Milk DNA can also be used to diagnose particular types of infections. Milk DNA purified using Norgen’s kit is of the highest quality, and is compatible with a number of downstream applications including PCR, Southern Blot analysis, sequencing and microarray analysis.

Milk DNA Preservative

Norgen’s Milk DNA Preservation Solution is an aqueous storage buffer designed for rapid cellular lysis and subsequent preservation of DNA from fresh specimens. The preservative prevents the growth of Gram-negative and Gram-positive bacteria and fungi, and also inactivates viruses allowing the resulting non-infectious samples to be handled and shipped safely. In addition, the preservative eliminates the need to immediately process or freeze samples and allows the samples to be shipped to centralized testing facilities at ambient temperature. The components of the preservative allow samples to be stored for 1 month without any detectable DNA degradation.

Details

Supporting Data

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Kit Specifications
Maximum Milk Input	0.5 mL
Time to Complete 10 Purifications	80 minutes (20 minutes hands on)

Storage Conditions and Product Stability
The Proteinase K should be stored at -20°C upon arrival and after reconstitution. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.

Storage in Preservation Buffer
1) Storage at -20°C is recommended for archival samples and will provide optimal preservation. The preservation buffer will freeze at -20°C. Samples can be stored indefinitely at -70°C.
2) Samples can be stored at room temperature (25°C) for up to 1 month without significant loss of DNA quality.
3) DNA has also been successfully isolated from samples stored at 37°C 1 for one week.

Kit Components	Cat. 44800 (25 preps)
Preservation Solution E	30 mL
Proteinase K	0.6 mL
Purification Additive	0.5 mL
Buffer SK	40 mL
Wash Solution A	18 mL
Elution Buffer B	8 mL
Mini Spin Columns	25
Collection Tubes	25
Elution Tubes (1.7 mL)	25
Product Insert	1

Sulfo DBCO-PEG4-Maleimide is a water-soluble click chemistry reagent with a maleimide group and a DBCO moiety. The hydrophilic PEG spacer arm improves solubility in aqueous buffers. Maleimide group specifically and efficiently reacts with thiols to form stable thioether bonds. The low mass weight will add minimal spacer to modified molecules and will enable simple and efficient incorporation of DBCO moiety onto cysteine-containing peptides or other thiol-containing biomolecules. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Sulfo DBCO-PEG4-Maleimide is a water-soluble click chemistry reagent with a maleimide group and a DBCO moiety. The hydrophilic PEG spacer arm improves solubility in aqueous buffers. Maleimide group specifically and efficiently reacts with thiols to form stable thioether bonds. The low mass weight will add minimal spacer to modified molecules and will enable simple and efficient incorporation of DBCO moiety onto cysteine-containing peptides or other thiol-containing biomolecules. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.