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.
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 |
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.
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:
Peelable heat sealing foil which is suitable for low temperature storage, high temperature uses and PCR.
These kits provide fast and reliable procedures for the purification of genomic DNA from viable yeast cells, fungal spores or mycelium, and bacteria including Gram-positive. Genomic DNA is efficiently extracted from the cells by a combination of heat treatment, detergents and Bead Tubes (provided). An optional lyticase treatment allows for improved DNA yields with certain fungal and yeast species. Recovered genomic DNA is of excellent yield and purity for any downstream application including PCR, qPCR, Restriction Fragment Length Polymorphism (RFLP), Amplified Fragment Length Polymorphism (AFLP), sequencing, SNP analysis and more.
Fungi/Yeast Genomic DNA Isolation Kit (Spin Column)
This kit provides rapid spin column purification of genomic DNA from viable yeast cells, fungal spores or mycelium, and bacteria including Gram-positive. Preparation time for a single sample is less then 30 minutes, and each kit contains sufficient materials for 50 preparations.
Fungi/Yeast Genomic DNA Isolation 96-Well Kit (HT)
Norgen’s Fungi/Yeast Genomic DNA Isolation 96-Well Kit provides a fast, reliable and simple procedure for high throughput isolation of DNA from viable yeast cells, fungal spores or mycelium and Gram-positive bacteria. The purification could be performed on either a vacuum manifold or using centrifugation. Complete 96 purifications in 40 minutes.
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| Kit Specifications | |
| Binding Capacity Per Well | 50 μg |
| Maximum Loading Volume Per Well | 500 μL |
| Size of DNA Purified | All sizes |
| Maximum Amount of Starting Material: Fungi (Wet weight) Yeast or Gram-positive bacterial culture | 50 mg 0.5 mL – 1 mL |
| Time to Complete 96 Purifications | 40 minutes |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 2 years in their unopened containers.
| Component | Cat. 27300 (50 preps) | Cat. 27350 (192 preps) |
|---|---|---|
| Lysis Buffer L | 30 mL | 2 x 60 mL |
| Resuspension Solution A | 20 mL | 60 mL |
| Solution BX | 28 mL | 2 x 28 mL |
| Wash Solution A | 18 mL | 2 x 38 mL |
| Elution Buffer B | 15 mL | 30 mL |
| Bead Tubes | 50 | 200 |
| Spin Columns | 50 | – |
| Collection Tubes | 50 | – |
| 96-Well Plate | – | 2 |
| 96-Well Collection Plate | – | 2 |
| Adhesive Tape | – | 4 |
| Elution Tubes (1.7 mL) | 50 | – |
| 96-Well Elution Plate | – | 2 |
| Product Insert | 1 | 1 |