Current Microbiology

, Volume 29, Issue 5, pp 301–307 | Cite as

An effective method to extract DNA from environmental samples for polymerase chain reaction amplification and DNA fingerprint analysis

  • L. Arlene Porteous
  • John L. Armstrong
  • Ramon J. Seidler
  • Lidia S. Watrud


A rapid direct-extraction method was used to obtain DNA from environmental soil samples. Heat, enzymes, and guanidine isothiocyanate were utilized to lyse cells. The DNA was purified by agarose gel electrophoresis, amplified with 16S rRNA-based primers by use of the polymerase chain reaction, and then digested with the restriction endonucleasePalI. The extraction method was used to obtain DNA from a variety of plants, bacteria, and fungi includingGossypium hirsucum (cotton),Pseudomonas, Bacillus, Streptomyces, andColletotrichum. Up to 100 μg DNA/g (wet weight) of soil and 400 μg DNA/g of plant material were recovered. Restriction endonuclease analysis patterns of amplified rDNA from pure microbial cultures and plant species contained three to five different DNA fragments. Amplified rDNA of mixed population DNA extracts from soil samples, digested with the restriction endonucleasePalI, contained 12–20 DNA fragments, appearing as sample “fingerprints.” Results from eight environmental soil samples that were analyzed suggest that the amplified rDNA fingerprints can be used to help characterize the genetic and biological diversity of the microbial populations in these samples.


Bacillus Pseudomonas Streptomyces Environmental Sample Isothiocyanate 
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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Copyright information

© Springer-Verlag New York Inc. 1994

Authors and Affiliations

  • L. Arlene Porteous
    • 1
  • John L. Armstrong
    • 1
  • Ramon J. Seidler
    • 1
  • Lidia S. Watrud
    • 1
  1. 1.Biotechnology ProgramUnited States Environmental Protection Agency, Environmental Research LaboratoryCorvallisUSA

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