Plasmid DNA Isolation and Visualization: Isolation and Characterization of Plasmids from Clinical Samples

Methods Mol Biol. 2020;2075:3-20. doi: 10.1007/978-1-4939-9877-7_1.


Plasmids are important in carrying antibiotic resistance and other genes between bacterial cells, and a number of methods can be employed to characterize plasmids from clinical isolates. Single colonies typically obtained as part of hospital workflow can undergo S1 nuclease treatment to linearize plasmids followed by pulsed-field gel electrophoresis to enable determination of the number and sizes of plasmids present. Hybridization of S1/PFGE gels can be used to associate replicon types and passenger genes, such as those conferring antibiotic resistance, with a particular plasmid band. Individual plasmids, obtained by conjugation or transformation, can be compared by gel electrophoresis following restriction digestion of plasmid DNA prepared by alkaline lysis methods, including using specialized kits.

Keywords: Clinical isolate; Hybridization; Plasmid; Pulsed-field gel electrophoresis; Restriction digest; S1 nuclease; Transformation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Conjugation, Genetic
  • Electrophoresis, Gel, Pulsed-Field / methods
  • Humans
  • Molecular Diagnostic Techniques* / methods
  • Nucleic Acid Hybridization / methods
  • Plasmids / genetics*
  • Plasmids / isolation & purification*
  • Transformation, Genetic