DNA fingerprinting of Vibrio cholerae and Aeromonas species by pulsed-field minigel electrophoresis.

Full 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.

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Author information

Author/s: Lopez-Canovas, Lilia (L); Bravo, Laura (L); Herrera, Jose (J); Riveron, Ana Maria (AM); Javer, Elisa (E); Sanchez, Axel (A); Fando, Rafael (R); Noa, Maria Dolores (MD); Fernandez, Anabel (A);

Affiliation: Department of Molecular Biology, Cuban Neuroscience Center, Havana, Cuba. llc(-atsign-)cneuro.edu.cu

Journal and publication information

Publication Type: Journal Article

Journal: Electrophoresis (Electrophoresis), published in Germany. (Language: eng)

Reference: 2006-Jul; vol 27 (issue 14) : pp 2857-64

Dates: Created 2006/08/23; Completed 2006/10/06;

PMID: 16721902, status: MEDLINE (last retrieval date: 12/26/2008)

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Aeromonas - genetics; isolation & purification
Bacterial Typing Techniques
DNA Fingerprinting

DNA, Bacterial - analysis; chemistry
Electrophoresis, Gel, Pulsed-Field - methods
Vibrio cholerae - genetics; isolation & purification

Associated Chemicals: DNA, Bacterial (0)

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