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DNA fingerprinting of Vibrio cholerae and Aeromonas species by pulsed‐field minigel electrophoresis
Author(s) -
LopezCanovas Lilia,
Bravo Laura,
Herrera Jose,
Riveron Ana Maria,
Javer Elisa,
Sanchez Axel,
Fando Rafael,
Noa Maria Dolores,
Fernandez Anabel
Publication year - 2006
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200500629
Subject(s) - vibrio cholerae , electrophoresis , gel electrophoresis , dna , aeromonas , dna profiling , chromatography , chemistry , resolution (logic) , biology , microbiology and biotechnology , bacteria , biochemistry , genetics , artificial intelligence , computer science
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.