Comparison of DNA Extraction and Purification Methods from Different Soils for Metagenomic Sequencing

Abstract:

There is an increased interest in the extraction of nucleic acids from various environmental samples, since only a minority of naturally occurring microbes can be cultured using standard techniques. Nucleic acids extraction and purification from soils are extremely challenging due to the low biomass, high organic contents and high variability of soil types. This has been regarded as one of the major difficulties that hamper the development of soil microbial ecology study. No commercial nucleic acids kits currently available are capable of preparing the DNAs without modifications. The cost can be very high for DNA extraction from extreme environmental soil samples, such as soils that have extreme high or low pHs. In this work, we developed and optimized soil DNA extraction and purification methods on different soils and compared the impact of three different DNA extraction protocols on DNA yield and purity. For the three different types of soil we used, direct extraction obtained the highest DNA recover rate, but required more cleanup steps. MoBio PowerSoil® DNA Isolation Kit yields less but do not require as many downstream cleaning steps. Both of the two methods obtained a more abundant microbial community than Meta-G-Nome^TM DNA Isolation Kit.

• Periodical: Advanced Materials Research (Volumes 955-959)
• Pages: 306-309
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.955-959.306
• Online since: June 2014
• Authors: Lin Hui Wu, Jian Li Liu, Jing Zeng, Ji Zhao*
• Keywords: DNA Extraction, DNA Purification, DNA Yield

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