Pulsed-Field Gel Electrophoresis (PFGE) Analysis of Listeria monocytogenes

Methods Mol Biol. 2021:2220:79-88. doi: 10.1007/978-1-0716-0982-8_6.

Abstract

PFGE is a valuable tool for assessing L. monocytogenes strain interrelatedness. It is based on the study of total bacterial DNA restriction patterns. Cells are embedded in agarose plugs before being lysed. The released DNA is then digested into large fragments by restriction enzymes. As DNA fragments are too large to be separated by traditional electrophoresis in an agarose gel, changes in the direction of the electrical current are periodically applied in order to allow the proper migration of large DNA fragments. Strains are characterized by the obtained DNA fragment patterns or pulsotypes which vary depending on the number and size of bands.

Keywords: Electrophoresis; Listeria monocytogenes; PFGE; Pulsotype; Restriction.

MeSH terms

  • DNA Restriction Enzymes / chemistry
  • DNA, Bacterial / analysis*
  • DNA, Bacterial / genetics
  • Electrophoresis, Gel, Pulsed-Field / methods*
  • Food Microbiology
  • Humans
  • Listeria monocytogenes / genetics
  • Listeria monocytogenes / isolation & purification*
  • Listeriosis / microbiology
  • Sepharose / chemistry

Substances

  • DNA, Bacterial
  • Sepharose
  • DNA Restriction Enzymes