Introduction

Soil samples contain a large number of microorganisms, the vast majority of which can not be directly cultivated for reproduction and research. Extracting DNA from soil samples is the most effective method for studying soil microorganisms. At present, there are mainly direct and indirect methods for extracting microbial DNA from soil samples. The direct method refers to placing soil samples in the lysis solution, and using effective wall breaking methods to release all microbial DNA into the lysis solution, followed by separation and extraction, such as Zhou’s method. Indirect method refers to placing soil in a buffer, such as Buffer PBS, to separate microorganisms from the soil and then extract DNA. The indirect method can greatly reduce the impact of humic acids and heavy metal salts on DNA extraction in soil, but this method will lose many microorganisms and the resulting DNA is not the entire genome (metagenome) of the soil sample. Currently, few researchers have adopted this method. Extracting DNA directly from soil samples can maximize the likelihood of obtaining the entire genome, but this method faces the following issues:

1. Humic acid pollution. The soil, especially in forests and grasslands, is rich in humic acids. Humic acid is a series of organic molecules, some of which are very similar to nucleic acid molecules and difficult to remove during purification. Trace amounts of humic acid pollution can lead to downstream applications such as PCR and enzyme digestion failure.

2. Lysis method. Soil samples contain various microorganisms, such as bacteria and fungi. Gram positive bacteria and fungi both contain very thick bacterial walls, and effectively breaking down the cell walls of these microorganisms is crucial for extracting high-yield metagenomic DNA. Due to the complexity of soil samples, it is not feasible to use enzymatic methods (such as lysozyme, wall breaking enzyme, snail enzyme) or liquid nitrogen grinding, as the soil contains various metalions or inhibitory factors that inactive the digestive enzymes, or the presence of sand particles in the soil makes liquid nitrogen grinding difficult.

3. The DNA yield is difficult to control. Soil samples would have significant changes in the number and variety of microorganisms due to fertility, inferiority, high moisture content, dryness, or depth of sampling. In a small range of soil samples, the DNA content often varies by thousands of times. In addition, certain chemical components in soil, such as heavy metal salts and clay substances, can cause a decrease in DNA yield.

Magen’s HiPure Soil DNA Kits are currently the most optimized kit for soil DNA extraction. The kit adopts glass bead grinding method and thermal shock chemical wall breaking method, which can be carried out in the point vortex instrument without special bead grinding instrument, and is suitable for a wide range of laboratories. The Absorber Solution in the reagent kit is a humic acid adsorbent exclusively developed by Magen Company, which can efficiently remove various humic acid pollutants. In addition, an alcohol-free silica gel column purification method is also used to efficiently remove various soluble metal salts and other soluble inhibitory factors from the soil. The kit has successfully extracted from the following soil (partially based on customer feedback): soil from forests in nature reserves (30 to 40 years old forest soil with a surface layer of 30-50cm deciduous layer), mangrove soil, grasslands, farmland, seabed mud, sludge, mineral area soil, organic matter contaminated soil, pond mud, garbage mud, air conditioning pipeline deposits, etc.

This product allows rapid and reliable isolation of high-quality genomic DNA from various soil samples. Up to 500 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 versatilityof 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 DNA from 200-500mg soil sample
Applications	PCR, southern blot and enzyme digestion, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Soil
Sample amount	200-500mg
Elution volume	≥30μl
Time per run	≤60 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

Soil 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

• Fast – several samples can be extracted in 40 minutes (after digestion)
• High purity – purified DNA can be directly used in various downstream applications
• Good repeatability – silica technology can obtain ideal results every time
• High recovery – DNA can be recovered at the level of PG

Kit Contents

Contents	D314202	D314203
Purification Times	50 Preps	250 Preps
Hipure DNA Mini Columns II	50	250
2ml Collection Tubes	50	250
2ml Bead Tubes	50	250
Buffer SOL	60 ml	250 ml
Buffer SDS	5 ml	20 ml
Buffer PS	10 ml	50 ml
Absorber Solution	10 ml	50 ml
Buffer GWP	40 ml	220  ml
Buffer DW1	30 ml	150  ml
Buffer GW2*	20 ml	2 x 50  ml
Buffer AE	15 ml	30 ml

Storage and Stability

Absorber Solution should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

Experiment Data

Soil samples contain a large number of microorganisms, the vast majority of which can not be directly cultivated for reproduction and research. Extracting DNA from soil samples is the most effective method for studying soil microorganisms. At present, there are mainly direct and indirect methods for extracting microbial DNA from soil samples. The direct method refers to placing soil samples in the lysis solution, and using effective wall breaking methods to release all microbial DNA into the lysis solution, followed by separation and extraction, such as Zhou’s method. Indirect method refers to placing soil in a buffer, such as Buffer PBS, to separate microorganisms from the soil and then extract DNA. The indirect method can greatly reduce the impact of humic acids and heavy metal salts on DNA extraction in soil, but this method will lose many microorganisms and the resulting DNA is not the entire genome (metagenome) of the soil sample. Currently, few researchers have adopted this method. Extracting DNA directly from soil samples can maximize the likelihood of obtaining the entire genome, but this method faces the following issues:

Introduction

This product provides fast and easy methods for purification of total DNA for reliable PCR and southern blotting. Total DNA (e.g., genomic, plasmid) can be purified from bacterial cultures.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from bacterial culture
Applications	PCR, southern blot and enzyme digestion, etc.
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Gram negative or positive bacteria
Sample amount	Bacterial culture: 0.5-1.5ml
Elution volume	≥60μl
Time per run	≤60 minutes

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Lysozyme. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• Fast – several samples can be extracted in 60 minutes (after digestion)
• High purity – unique adsorbent can completely remove inhibitory factors
• High recovery – DNA can be recovered at the level of PG

Kit Contents

Contents	D513101	D513102	D513103
Purification Times	48 Preps	96 Preps	480 Preps
MagPure Particles	1.2 ml	2.5 ml	11 ml
Lysozyme	60 mg	120 mg	600 mg
Proteinase K	24 mg	50 mg	220 mg
Protease Dissolve Buffer	3 ml	6 ml	30 ml
Buffer SDS	1.5 ml	3 ml	15 ml
Buffer MLA	30 ml	60 ml	300 ml
Buffer GW1*	13 ml	44 ml	110 ml
Buffer TE	30 ml	40 ml	200 ml

Storage and Stability

MagPure Particles, Lysozyme and Proteinase K 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 dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

This product provides fast and easy methods for purification of total DNA for reliable PCR and southern blotting. Total DNA (e.g., genomic, plasmid) can be purified from bacterial cultures.

Propargyl-PEG17-methane has an alkyne group which can participate in Click Chemistry reactions with azide compounds to yield a stable triazole linkage; copper is required as a catalyst. The hydrophilic PEG units increases solubility of the molecules in aqueous environment. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG17-methane has an alkyne group which can participate in Click Chemistry reactions with azide compounds to yield a stable triazole linkage; copper is required as a catalyst. The hydrophilic PEG units increases solubility of the molecules in aqueous environment. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.