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DNA isolation from forest soil suitable for PCR assays of fungal and plant rRNA genes

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Abstract

This protocol for DNA isolation from forest soil samples is advantageous because it uses only one liquid transference step and can process several samples with minimal time and equipment. The use of benzyl chloride early in the extraction protocol increases DNA yield and purity. The obtained DNA is useful for PCR amplification of nuclear and mitochondrial ribosomal related sequences from fungi and ribosomal DNA from plant chloroplasts. Isolated DNA can be used either undiluted or at low dilutions in PCR assays. A final glassmilk treatment of isolated DNA is useful to recover high molecular weight DNA fractions from agarose gel. DNA losses during glassmilk treatment can generate negative PCR results.

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Abbreviations

BC:

benzyl chloride

ITS:

internal transcribed spacer of nuclear ribosomal unit

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Author notes

  1. Vázquez-Garciduenas

    Present address: Facultad de Medicina “Dr Ignacio Chávez”, División de Estudios de Posgrado, Universidad Michoacana de San Nicolás de Hidalgo, San Nicolas, Mexico

Authors and Affiliations

  1. Departamento de Bioquímica y Biotecnología de Plantas, CINVESTAV-IPN, Unidad Irapuato, Km 9.6, Libramiento Norte, Carretera Irapuato-León, A.P. 629, C.P. 36500, Irapuato, Gto., México

    Gerardo Vázquez-Marrufo, MA. Soledad,  Vázquez-Garciduenas, Blanca E. Gómez-Luna & Víctor Olalde-Portugal

Authors
  1. Gerardo Vázquez-Marrufo
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  2. MA. Soledad
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  3. Vázquez-Garciduenas
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  4. Blanca E. Gómez-Luna
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  5. Víctor Olalde-Portugal
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Vázquez-Marrufo, G., Soledad, M., Vázquez-Garciduenas et al. DNA isolation from forest soil suitable for PCR assays of fungal and plant rRNA genes. Plant Mol Biol Rep 20, 379–390 (2002). https://doi.org/10.1007/BF02772125

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