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DNA Isolation from Soil and Other Environmental Samples Made Easy with Patented Inhibitor Removal Technology (IRT)
Sponsored,vendor-submitted protocol   Sponsored by Mo Bio Laboratories, Inc.    Published in BioTechniques Protocol Guide 2009 (p.51) DOI: 10.2144/000113001

When processing soil and other environmental samples for DNA, you need a method that is fast, easy, and reliable. Meet your expectations with the PowerSoil™ DNA Isolation Kit from MO BIO Laboratories, Inc. PowerSoil™ is a key tool for studying microbial diversity, microbial detection, and applied genomics. When efficient cell lysis, high yields, and DNA purity are the most important considerations for reliable downstream applications, the PowerSoil™ DNA Isolation Kit is the method of choice by research institutes worldwide. Our patented technology removes PCR inhibitors from even the most challenging samples including those high in humic acid like soil, manure, compost, marine sediment and fresh or salt water. Pure eluted DNA ensures reliable PCR, which leads to more sensitive detection.

Introduction

MO BIO's PowerSoil™ DNA Isolation Kits are the only products which include patented Inhibitor Removal Technology (IRT). This new technology easily removes all PCR inhibitors including humic acids, fulvic acids, polysaccharides, polyphenolics, melanin, and hematin. Inhibitors are removed easily as high-quality DNA is isolated from different samples. The protocol involves a bead beating cell lysis step followed by the inhibitor removal step (IRT) where PCR inhibitors are selectively precipitated away from the desired genomic DNA. Finally, the DNA is purified using silica spin filter columns—the gold standard for DNA purification. Materials

PowerSoil™ DNA Isolation Kit
(Cat# 12888-50)

Vortex-Genie® 2 (Cat# 13111-V)

Vortex Adapters for Vortex-Genie® 2 holds12 (1.5–2.0 ml) tubes
(Cat# 13000-V1) or 24 (1.5–2.0 ml) tubes
(Cat# 13000-V1-24) Methods Lysis

1. Add 0.25 gm of soil to a PowerBead tube containing homogenization buffer.

2. Vortex 10 min using the vortex adapter on the Vortex-Genie® 2. Inhibitor Removal Technology

3. Add Solution C2 and incubate 5 min at 4°C. Centrifuge to remove inhibitors.

4. Add Solution C3 (IRT) and incubate 5 min at 4°C. Centrifuge to remove remaining inhibitors. Binding and Elution of DNA

5. Add Solution C4 to bind the DNA to the silica filter membrane spin column using centrifugation.

6. Wash the Spin Filter to remove salts.

7. Elute DNA in 10 mM Tris and use in PCR or qPCR assays. Results

DNA was isolated from nine different sources of soil ranging in degree of PCR inhibitor levels. Compared with two other suppliers' products for soil DNA extraction, the PowerSoil™ DNA Isolation Kit provided the most consistent results, highest yields, and best integrity of DNA (Figure 1).


Figure 1. Genomic DNA from environmental samples. (Click to enlarge)


PCR amplification of genomic DNA purified in Figure 1 using eubacterial primers was performed using 1 µl of the eluates. Only the PowerSoil™ DNA Isolation Kit prepared DNA was successful in all PCR amplifications while the competitor products were not sufficiently pure to allow for PCR amplification in all cases (Figure 2).


Figure 2. PCR amplification of genomic DNA. (Click to enlarge)


Typical measures of purity are the A260/280 and A260/230 readings where high purity is indicated by a ratio of 1.8-2.0 and >2.0, respectively. Results using the NanoDrop® spectrophotometer for DNA purified using the PowerSoil™ DNA Soil Isolation Kit are shown in Table 1. These readings are reliable indicators that the DNA will be good substrates for downstream applications.


Table 1. (Click to enlarge)


Trust the world leader for soil DNA and RNA isolation when processing your rare and irreplaceable samples!

Correspondence

Address correspondence to MO BIO Laboratories, Inc., 2746 Loker Ave. West, Carlsbad, CA, 92010 USA; Tel.: (800) 606-6246; Fax: (760) 929-0109; [email protected], www.mobio.com.

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