Name: Soil DNA Isolation Plus Kit
Brand: Norgen Biotek Corp.
SKU: 64000
Price: 315.000000 USD
Availability: InStock

Features and Benefits

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.

Kit Specifications
Maximum Soil Input	250 mg
Type of Soil Processed	All soil types
Maximum Column Binding Capacity	50 μg
Maximum Column Loading Volume	650 μL
Time to Complete 10 Purifications	30 minutes

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

PROTOCOLS

(64000) Soil DNA Isolation Plus Kit - Protocol (50 prep)

APPLICATION NOTES

The Effect of Sample Preparation on Humic Acid Removal from Peat Samples: A Comparative Study

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

Why do I have poor DNA recovery?

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.

Why is the purified DNA not performing well in downstream applications?

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	Metagenomic Influential Insights in the Formation of Biogenic Iron Hydroxysulfate Precipitates by Ferrous Oxidative Microbial Consortia
Citation	Geomicrobiology Journal 2023.
Authors	Daladier M. Castilo, Jesus P. Medina, Francisco Gamarra, Sandra Delgado, Virginia Chipana, Claudia Clavijo, Robert Ccorahua-Santo and Mesalina Peceros-Melchor

Title	Molecular identification of Acanthamoeba spp., Balamuthia mandrillaris and Naegleria fowleri in soil samples using quantitative real-time PCR assay in Turkey; Hidden danger in the soil!
Citation	Acta Tropica 2023.
Authors	Mehmet Aykur a b, Hande Dagci b

Title	Startup performance and microbial communities of a decentralized anaerobic digestion of food waste
Citation	Chemosphere 2023.
Authors	Yong Wei Tiong, Pooja Sharma, Hailin Tian, To-Hung Tsui, Heng Thong Lam, Yen Wah Tong,

Title	Systematic screening of carbon-based anode materials for bioelectrochemical systems
Citation	Journal of Chemical Technology & Biotechnology 2023.
Authors	Zainab Ul, Pilar Sánchez-Peña, Mireia Baeza, Mira Sulonen, David Gabriel, Juan Antonio Baeza, Albert Guisasola

Title	A Comparison of Performance between Two Anaerobic Biodigesters Configurations for Biogas Production.
Citation	Revista Técnica Energía 2018.
Authors	Ramirez-Perez, J. C. (2018)

Soil DNA Isolation Plus Kit