title: DNA fingerprinting of Vibrio cholerae and Aeromonas species by pulsed-field minigel electrophoresis.
abstract: DNA molecules of Vibrio cholerae and Aeromonas species were prepared by incubating immobilized cells for 4 and 2 h, respectively, with a nonenzymatic solution that contains chemical reagents only (NDSUPlus). This method gave results as reproducible as the enzymatic one that uses proteinase K, and rendered DNA molecules suitable for fingerprinting by mini-CHEF electrophoresis. As rapid DNA separations at high electric field are achieved in mini-CHEF chamber with low heat evolution, DNA restriction fragments were separated in 5 h at 10 V/cm in a single resolution window. Then, fragment separations in three resolution windows were done in 15 h. This time is shorter than the one needed by the large CHEF chamber for resolving fragments in a single resolution window. Three windows permitted to include larger numbers of restriction fragments in the calculation of isolate similarities. Both sample preparation and mini-CHEF electrophoresis may represent an alternative for performing massive epidemiological studies of V. cholerae and Aeromonas species.
authors.affiliation: Department of Molecular Biology, Cuban Neuroscience Center, Havana, Cuba. llc@cneuro.edu.cu
authors.forename: Lilia
authors.initials: L
authors.lastname: Lopez-Canovas
authors.forename: Laura
authors.initials: L
authors.lastname: Bravo
authors.forename: Jose
authors.initials: J
authors.lastname: Herrera
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Riveron
authors.forename: Elisa
authors.initials: E
authors.lastname: Javer
authors.forename: Axel
authors.initials: A
authors.lastname: Sanchez
authors.forename: Rafael
authors.initials: R
authors.lastname: Fando
authors.forename: Maria Dolores
authors.initials: MD
authors.lastname: Noa
authors.forename: Anabel
authors.initials: A
authors.lastname: Fernandez
mesh_terms: D000333:Aeromonas / Q000235:genetics / Q000302:isolation & purification*; D015373:Bacterial Typing Techniques; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000032:analysis* / Q000737:chemistry; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D014734:Vibrio cholerae / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D004269:DNA, Bacterial