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A simple mini-method to extract DNA directly from soil for use with polymerase chain reaction amplification

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Abstract

A rapid, simple method is used that yields amplifiable fungal and bacterial DNAs directly from soil. DNA is separated from soil contaminants by electrophoresis in low-melting-temperature agarose and used directly in polymerase chain reaction amplification. Fifty 20-mg samples can be processed in one day. Fragments of 16S and 18S ribosomal RNAs are amplified by polymerase chain reaction with DNA extracted from the soil. Universal primers are used that are capable of amplifying ribosomal DNA from a wide variety of bacteria and fungi. Eubacterial and fungal primers are used that are capable of distinguishing between eubacterial and fungal DNAs. Restriction enzyme digests are performed on amplified DNA fragments from five soil samples.

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Literature Cited

  1. Edwards K, Johnstone C, Thompson C (1991) A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucleic Acids Res 19:1349

    PubMed  Google Scholar 

  2. Giovannoni SJ, Britschgi TB, Moyer CL, Field KG (1990) Genetic diversity in Sargasso Sea bacterioplankton. Nature 344:60–63

    PubMed  Google Scholar 

  3. Neefs J-M, Van de Peer Y, Hendriks L, De Wachter R (1990) Compilation of small ribosomal subunit RNA sequences. Nucleic Acids Res 18:2237–2330

    PubMed  Google Scholar 

  4. Porteous LA, Armstrong JL (1991) Recovery of bulk DNA from soil by a rapid, small-scale extraction method. Curr Microbiol 22:345–348

    Google Scholar 

  5. Rochelle PA, Olson BH (1991) A simple technique for electro-elution of DNA from environmental samples. Bio Techniques 11:724–728

    Google Scholar 

  6. Schmidt TM, DeLong EF, Pace NR (1991) Analysis of a marine picoplankton community by 16S rRNA gene cloning and sequencing. J Bacteriol 173:4371–4378

    PubMed  Google Scholar 

  7. Steffan RJ, Atlas RM (1991) Polymerase chain reaction: applications in environmental microbiology. Annu Rev Microbiol 45:137–161

    PubMed  Google Scholar 

  8. White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. New York: Academic Press, pp 315–322

    Google Scholar 

  9. Zintz CB, Beebe DC (1991) Rapid reamplification of PCR products purified in low melting point agarose gels. Bio Techniques 11:158–162

    Google Scholar 

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Author notes

  1. John L. Armstrong

    Present address: College of Education, Oregon State University, 97331, Corvallis, OR, USA

Authors and Affiliations

  1. Biotechnology Program, United States Environmental Protection Agency, Environmental Research Laboratory, 200 S.W. 35th St., 97333, Corvallis, Oregon, USA

    L. Arlene Porteous & John L. Armstrong

Authors
  1. L. Arlene Porteous
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  2. John L. Armstrong
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Porteous, L.A., Armstrong, J.L. A simple mini-method to extract DNA directly from soil for use with polymerase chain reaction amplification. Current Microbiology 27, 115–118 (1993). https://doi.org/10.1007/BF01570868

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  • DOI: https://doi.org/10.1007/BF01570868

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