Pulsed-field electrophoresis in contour-clamped homogenous electric fields (CHEF) for fingerprinting of Rhizobium spp |
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Authors: | Viviana Corich Alessio Giacomini Francisco J. Ollero rea Squartini Marco F. Nuti |
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Affiliation: | Dipartimento di Biotecnologie Agrarie, Universitàdi Padova, Padova, Italy |
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Abstract: | Abstract: A pulsed-field gel electrophoretic method based on contour-clamped homogeneous electric field (CHEF) was developed for the analysis of natural isolates of Rhizobium leguminosarum biovar viciae . The procedure involves the use of 'rare-cutting' endonucleases. The separation of genomic DNA fragments with split runs ( τ = 5 s for 15 h followed by τ = 12 s for 9 h) allows a clear definition of profiles with bands ranging from 20–300-kb pairs. Two and a half days are sufficient to reproducibly accomplish the procedure from cell lysis to gel picture. The method can be used for different fast-growing rhizobia. |
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Keywords: | Pulsed-field electrophoresis Rhizobium Fingerprinting |
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