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Open Access Methodology article

Updated 16S rRNA-RFLP method for the identification of all currently characterised Arcobacter spp

María José Figueras1*, Arturo Levican1 and Luis Collado2

Author Affiliations

1 Unitat de Microbiologia, Departament de Ciències Mediques Bàsiques, Facultat de Medicina i Ciències de la Salut. IISPV, Universitat Rovira i Virgili, Reus, Spain

2 Institute of Biochemistry and Microbiology, Faculty of Sciences, Universidad Austral de Chile, Valdivia, Chile

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BMC Microbiology 2012, 12:292  doi:10.1186/1471-2180-12-292

Published: 18 December 2012

Additional files

Additional file 1:

Table S1. Computer simulated profiles of Arcobacter spp. 16S rRNA gene (1026 bp) digestion with MseI endonuclease. Species with specific RFLP patterns are in bold.

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Additional file 4:

Figure S1. Microheterogeneities (or mutations) in the 16S rRNA gene of seven atypical A. cryaerophilus strains in relation to the type strain (LMG 9904T), strain LMG 10829 (A. cryaerophilus subgroup 1B) and the type strain ofA. butzleri (LMG 10828T). Sequence alignment of the 16S rRNA gene (positions 190–207 in relation to Escherichia coli) of seven atypical A. cryaerophilus strains showing mutations at positions 192 (T→C) and 205 (A→G), which alter the MseI restriction enzyme recognition site (TTAA). IUPAC code, Y = Pyrimidine (C or T); R = Purine (A or G).

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Additional file 5:

Figure S2. Agarose gel (3.5%) comparing the 16S rRNA-RFLP patterns obtained using endonucleases a\) TasI and b) MnlI for species A. butzleri,A. thereius and A. trophiarum. Lanes 1 and 14, 50 bp ladder (Fermentas); 2, A. butzleri LMG 10828T; 3, A. butzleri F42; 4, A. butzleri F43; 5, A. butzleri F44; 6, A. butzleri F50; 7, A. butzleri LMG 11118; 8, A. thereius LMG 24486T; 9, A. thereius SW24; 10, A. thereius F89-4; 11, A.thereius F93-4 y 12, A.thereius LMG 24487; 13, A. trophiarum CECT 7650 (identical pattern to that of the 11 atypical strains of A. cryaerophilus, Additional file 2: Table S2). MnlI was selected because it produced more distinctive patterns among the species than TasI.

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Additional file 2:

Table S2. Computer simulated profiles of Arcobacter spp.16S rRNA gene (1026 bp) digestion with MnlI endonuclease. Species in bold are those that show a specific RFLP pattern that was not distinguished with MseI.

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Additional file 3:

Table S3. Computer simulated profiles of Arcobacter spp. 16S rRNA gene (1026 bp) digestion with BfaI endonuclease. Species in bold are those that now show a specific RFLP pattern that was not distinguished previously with MseI or MnlI.

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