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      Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA

      1 , 1 , 1
      Applied and Environmental Microbiology
      American Society for Microbiology

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          Abstract

          We describe a new molecular approach to analyzing the genetic diversity of complex microbial populations. This technique is based on the separation of polymerase chain reaction-amplified fragments of genes coding for 16S rRNA, all the same length, by denaturing gradient gel electrophoresis (DGGE). DGGE analysis of different microbial communities demonstrated the presence of up to 10 distinguishable bands in the separation pattern, which were most likely derived from as many different species constituting these populations, and thereby generated a DGGE profile of the populations. We showed that it is possible to identify constituents which represent only 1% of the total population. With an oligonucleotide probe specific for the V3 region of 16S rRNA of sulfate-reducing bacteria, particular DNA fragments from some of the microbial populations could be identified by hybridization analysis. Analysis of the genomic DNA from a bacterial biofilm grown under aerobic conditions suggests that sulfate-reducing bacteria, despite their anaerobicity, were present in this environment. The results we obtained demonstrate that this technique will contribute to our understanding of the genetic diversity of uncharacterized microbial populations.

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          Author and article information

          Journal
          Applied and Environmental Microbiology
          Appl Environ Microbiol
          American Society for Microbiology
          0099-2240
          1098-5336
          March 1993
          March 1993
          : 59
          : 3
          : 695-700
          Affiliations
          [1 ]Department of Chemistry, Leiden University, The Netherlands.
          Article
          10.1128/aem.59.3.695-700.1993
          202176
          7683183
          ff00d071-2d3c-4150-84f6-72f9c1f7e4b3
          © 1993

          https://journals.asm.org/non-commercial-tdm-license

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