Soil DNA Isolation Plus Kit
For the detection of microorganisms from soil samples
For research use only and NOT intended for in vitro diagnostics.
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For the detection of microorganisms from soil samples
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Overview
- Rapid and convenient method to detect microorganisms in soil samples
- Process all soil types
- Remove organic substances using the OSR Solution
- Remove all humic acid from DNA samples
- Fast and easy processing using a rapid spin-column format
- Isolate high quality total DNA from a variety of microorganisms including bacteria, fungi and algae
Norgen's Soil DNA Isolation Plus Kit provides a convenient and rapid method for the detection of microorganisms from soil samples. All types of soil samples can be processed with this kit, including common soil samples and difficult soil samples with high humic acid content such as compost and manure. The kit removes all traces of humic acid and PCR inhibitors using the provided the OSR (Organic Substance Removal) Solution. A simple and rapid spin column procedure is then used to further purify the DNA. Total genomic DNA can be isolated and purified from all the various microorganisms found in soil, such as bacteria, fungi and algae. The purified DNA is of the highest quality and is fully compatible with downstream PCR applications, as all humic acid substances and PCR inhibitors are removed during the isolation.
Details
Supporting Data
Figure 1. Comparison of DNA Yield from Top Soil and Clay Samples. Norgen’s Soil DNA Isolation Plus Kit (Cat. 64000) and Competitor M’s kit were used to isolate DNA from 250 mg of top soil and clay samples. Following isolation, 10 µL from each 100 µL elution was loaded on 1% TAE agarose gel. Lane M: Norgen's HighRanger 1kb DNA Ladder.
Figure 2. Comparison of DNA Concentration Isolated from Top Soil and Clay Samples. Norgen’s Soil DNA Isolation Plus Kit (Cat. 64000) and Competitor M’s kit were used to isolate DNA from 250 mg top soil and clay samples. Norgen’s kit showed higher DNA concentrations for both samples in comparison to the competitor kit.
Figure 3. High Quality DNA confirmed by Real-time PCR. Soil DNA was isolated from 250 mg of clay (containing low humic acids) and top soil (containing high humic acids) samples using Norgen's Soil DNA Isolation Plus Kit and Competitor M's Kit. DNA quality was confirmed by Real-time PCR using 4 µL of soil DNA (total PCR reaction volume was 20 µL) to detect 16s rDNA from different soil samples. The earlier Ct value with Norgen's DNA samples (red lines) compared to Competitor M's samples (green lines) indicated a higher quality of soil DNA for downstream applications.
Figure 4. Microbial profiles from soil samples (clay and top soil) isolated using Norgen’s Soil DNA Isolation Plus Kit (Cat. 64000) and Competitor M's kit. The relative abundance of phylum-level classifications indicates the efficiency of DNA isolation and the qualityof DNA. 16S rRNA genes (v3-4 region) were amplified and the amplicons were sequenced on Illumina® MiSeq™ (MiSeq Reagent Kit v3) using Norgen’s 16S Metagenomic workflow with Illumina 16S Metagenomics Pipeline (v1.0.1).
Figure 5. Abundance of bacterial genera heat map based on phylum level between two DNA soil samples isolation methods; Norgen’s Soil DNA Isolation Plus Kit (Cat. 64000) and Competitor M's kit. The numbers indicate species count.
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Kit Specifications
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| Maximum Soil Input |
250 mg
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| Type of Soil Processed |
All soil types
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| Maximum Column Binding Capacity |
50 μg
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| Maximum Column Loading Volume |
650 μL
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| Time to Complete 10 Purifications |
30 minutes
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Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year from the date of shipment.
Component | Cat. 64000 (50 preps) |
|---|---|
Lysis Buffer D | 45 mL |
Lysis Additive A | 2 x 6 mL |
Binding Buffer I | 7 mL |
OSR Solution | 3 mL |
Lysis Buffer QP | 25 mL |
Wash Solution A | 18 mL |
Elution Buffer B | 8 mL |
Bead B Tubes | 50 |
Spin Columns | 50 |
Collection Tubes | 50 |
Elution Tubes (1.7 mL) | 50 |
Product Insert | 1 |
Documentation
Norgen’s Soil DNA Isolation Kit Outperforms a Competitor with Effective Humic Acid Removal for Metagenomic Studies
Norgen’s Improved Soil DNA Isolation Plus Kit Produces Yields and Quality Comparable With a Top Competitor
FAQs
Plus
Poor DNA recovery could be due to one or more of the following:
- Homogenization was incomplete.
Depending on the type of soil, further vortexing with the flat bed vortex or bead beater equipment may be required. However, it is not recommended to increase the vortex time to longer than 10 minutes at maximum speed.
- Lysis Additive A was not added to the lysate.
Ensure that the provided Lysis Additive A is added to separate humic acid and increase DNA yield.
- Lysis Buffer QP and Ethanol were not added to the lysate.
Ensure that 400 µL of Lysis Buffer QP and 550 µL of 96-100% ethanol are added to the lysate before binding to the column.
- Ethanol was not added to the Wash Solution A.
Ensure that 42 mL of 96-100% ethanol is added to the supplied Wash Solution A prior to use.
If the DNA does not perform well in downstream applications, it may be due to one or more of the following:
- Eluted DNA sample is brown.
The elution contains high humic acids. Ensure that the OSR Solution was added to the clean lysate. Also, ensure the column was washed with Binding Buffer B.
- DNA was not washed with the provided Binding Buffer B and Wash Solution A.
Traces of humic acids or salt from the binding step may remain in the sample if the column is not washed with the provided Binding Buffer B and Wash Solution A. Humic acids and salt may interfere with downstream applications, and thus must be washed from the column.
- Ethanol carryover.
Ensure that the dry spin under the Column Wash procedure is performed in order to remove traces of ethanol prior to elution. Ethanol is known to interfere with many downstream applications.
- PCR reaction conditions need to be optimized.
Take steps to optimize the PCR conditions being used, including varying the amount of template (10 ng to 50 ng for 20 µL of PCR reaction is recommended), changing the source of Taq polymerase, looking into the primer design, and adjusting the annealing conditions.
Citations
| Title | Enhancing bioelectrochemical hydrogen production from industrial wastewater using Ni-foam cathodes in a microbial electrolysis cell pilot plant |
| Journal | Water Research. 2024. |
| Authors | Oscar Guerrero-Sodric, Juan Antonio Baeza, Albert Guisasola |
| Title | Coupling anammox and heterotrophic denitrification activity at mainstream conditions in a single reactor unit |
| Journal | Chemical Engineering Journal. 2022. |
| Authors | Xènia Juan-Díaz, Lluc Olmo, Julio Pérez, Julián Carrera |
| Title | Comparative Metagenomics of Anaerobic Digester Communities Reveals Sulfidogenic and Methanogenic Microbial Subgroups in Conventional and Plug Flow Residential Septic Tank Systems |
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| Title | Ceramic-supported graphene oxide membrane bioreactor for the anaerobic decolorization of azo dyes |
| Journal | Journal of Water Process Engineering. 2022. |
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| Title | Evaluating the production and bio-stimulating effect of 5-methyl 1, hydroxy phenazine on microbial fuel cell performance |
| Journal | International Journal of Environmental Science and Technology. 2017. |
| Authors | S. Yousaf, M. Anam, N. Ali |
| Title | Comparison of Total DNA Extraction Methods for Microbial Community from Polluted Soil |
| Journal | Agriculture and Agricultural Science Procedia. 2015. |
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| Journal | Annals of Microbiology. 2015. |
| Authors | Giulio Moretti, Federica Matteucci, Claudia Ercole, Francesco Vegliò, Maddalena Del Gallo |
| Title | It takes an individual plant to raise a community: TRFLP analysis of the rhizosphere microbial community of two pairs of high- and low-metal-accumulating plants. |
| Journal | Soil Biology and Biochemistry. 2015. |
| Authors | Melanie P. Columbus, Sheila M. Macfie. |
| Title | Molecular Characterization and Potential of Bacterial Species Associated with Cassava Waste |
| Journal | Nigerian Food Journal. 2014. |
| Authors | A.I. Elijah, , O.O. Atanda, A.R. Popoola, S.V.A. Uzochukwu |
| Title | Insertion/Deletion-Based Approach for the Detection of Escherichia coli O157:H7 in Freshwater Environments. |
| Journal | American Chemical Society. 2014. |
| Authors | Shirley Y. Wong, Athanasios Paschos, Radhey S. Gupta, and Herb E. Schellhorn. |
| Title | Network topology reveals high connectance levels and few key microbial genera within soils. |
| Journal | Frontiers in Environmental Science. 2014. |
| Authors | Lupatini M, Suleiman A, Jacques R, Antoniolli Z, Ferreira A, Kuramae E and Roesch L. |
| Title | Optimization and Validation of Molecular Assays for Invasive Tunicate Monitoring in Environmental Water Samples. |
| Journal | Macrothink Institute. 2013. |
| Authors | Stewart-Clark S, Davidson J, Greenwood S. |
| Title | Bioremediation of high organic load lagoon sediments: Compost addition and priming effects. |
| Journal | Chemosphere. 2012. |
| Authors | d'Errico G, Giovannelli D, Montano C, Milanovic V, Ciani M, Manini E. |
| Title | Molecular Approaches to the Study of Biological Phosphorus Cycling. |
| Journal | Soil Biology. 2010. |
| Authors | Wasaki J. and Maruyama H. |
