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      Rapid procedure for detection and isolation of large and small plasmids.

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      Journal of bacteriology
      American Society for Microbiology

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          Abstract

          Procedures are described for the detection and isolation of plasmids of various sizes (2.6 to 350 megadaltons) that are harbored in species of Agrobacterium, Rhizobium, Escherichia, Salmonella, Erwinia, Pseudomonas, and Xanthomonas. The method utilized the molecular characteristics of covalently closed circular deoxyribonucleic acid (DNA) that is released from cells under conditions that denature chromosomal DNA by using alkaline sodium dodecyl sulfate (pH 12.6) at elevated temperatures. Proteins and cell debris were removed by extraction with phenol-chloroform. Under these conditions chromosomal DNA concentrations were reduced or eliminated. The clarified extract was used directly for electrophoretic analysis. These procedures also permitted the selective isolation of plasmid DNA that can be used directly in nick translation, restriction endonuclease analysis, transformation, and DNA cloning experiments.

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          Author and article information

          Journal
          J Bacteriol
          Journal of bacteriology
          American Society for Microbiology
          0021-9193
          0021-9193
          Mar 1981
          : 145
          : 3
          Article
          10.1128/jb.145.3.1365-1373.1981
          217141
          7009583
          7b28e9df-53d3-4664-aacd-2f72ff927455
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