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Soil contains compounds that inhibit PCR, so obtaining a metagenomic DNA isolation method that is free of these compounds is essential for the success of any metagenomic study that detects and explores the diversity of microorganisms associated with rhizosphere soils. This protocol describes an improved method for isolating high-quality, high-molecular-weight metagenomic DNA from rhizospheric soils. This protocol combines enzymatic, chemical, and physical strategies to ensure efficient cellular lysis and removal of humic contaminants that inhibit PCR. This method provides a high yield of good-quality metagenomic DNA from rhizospheric soils, which can be confirmed by spectrophotometric data and by amplification of the 16S rRNA gene by PCR.

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Isolation of Metagenomic DNA from Rhizosphere Soil

  • 章节

  • 发表日期：

    2025年

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  • Janet Jan-Roblero,
  • Hugo A. Hernández-Alvarez,
  • Juan A. Cruz-Maya

摘要

Soil contains compounds that inhibit PCR, so obtaining a metagenomic DNA isolation method that is free of these compounds is essential for the success of any metagenomic study that detects and explores the diversity of microorganisms associated with rhizosphere soils. This protocol describes an improved method for isolating high-quality, high-molecular-weight metagenomic DNA from rhizospheric soils. This protocol combines enzymatic, chemical, and physical strategies to ensure efficient cellular lysis and removal of humic contaminants that inhibit PCR. This method provides a high yield of good-quality metagenomic DNA from rhizospheric soils, which can be confirmed by spectrophotometric data and by amplification of the 16S rRNA gene by PCR.