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Current Microbiology

, Volume 27, Issue 2, pp 115–118 | Cite as

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

  • L. Arlene Porteous
  • John L. Armstrong
Article

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.

Keywords

Enzyme Polymerase Chain Reaction Agarose Soil Sample Electrophoresis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Literature Cited

  1. 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 19:1349PubMedGoogle Scholar
  2. 2.
    Giovannoni SJ, Britschgi TB, Moyer CL, Field KG (1990) Genetic diversity in Sargasso Sea bacterioplankton. Nature 344:60–63PubMedGoogle Scholar
  3. 3.
    Neefs J-M, Van de Peer Y, Hendriks L, De Wachter R (1990) Compilation of small ribosomal subunit RNA sequences. Nucleic Acids Res 18:2237–2330PubMedGoogle Scholar
  4. 4.
    Porteous LA, Armstrong JL (1991) Recovery of bulk DNA from soil by a rapid, small-scale extraction method. Curr Microbiol 22:345–348Google Scholar
  5. 5.
    Rochelle PA, Olson BH (1991) A simple technique for electro-elution of DNA from environmental samples. Bio Techniques 11:724–728Google Scholar
  6. 6.
    Schmidt TM, DeLong EF, Pace NR (1991) Analysis of a marine picoplankton community by 16S rRNA gene cloning and sequencing. J Bacteriol 173:4371–4378PubMedGoogle Scholar
  7. 7.
    Steffan RJ, Atlas RM (1991) Polymerase chain reaction: applications in environmental microbiology. Annu Rev Microbiol 45:137–161PubMedGoogle Scholar
  8. 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 315–322Google Scholar
  9. 9.
    Zintz CB, Beebe DC (1991) Rapid reamplification of PCR products purified in low melting point agarose gels. Bio Techniques 11:158–162Google Scholar

Copyright information

© Springer-Verlag New York Inc 1993

Authors and Affiliations

  • L. Arlene Porteous
    • 1
  • John L. Armstrong
    • 1
  1. 1.Biotechnology Program, United States Environmental Protection AgencyEnvironmental Research LaboratoryCorvallisUSA

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