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RpoB-PCR amplified gene and temporal temperature gradient gel electrophoresis: a rapid tool to analyse bacterial strains representative of cold-smoked salmon microflora ArchiMer
Giacomazzi, Sophie; Leroi, Francoise; L'Henaff, Cecile; Joffraud, Jean-jacques.
Aims: To evaluate rpoB gene as a biomarker of microbial biodiversity associated to cold-smoked salmon by a novel nested-polymerase chain reaction/temporal temperature gradient gel electrophoresis (PCR/TTGE) technique applied on pure cultures of reference strains. Methods and Results: DNA obtained from pure cultures of reference strains was used in a succession of a first PCR amplification of rpoB fragment with degenerated nonclamped primers and a nested-PCR with nondegenerated clamped primers. PCR products were then applied on a TTGE gel in order to analyse strains profile. High quantity of nested-PCR products were obtained for each tested strain and TTGE profiles showed a good separation between the different reference bacteria and an easy way to...
Tipo: Text Palavras-chave: TTGE; RpoB; Nested PCR; Cold smoked salmon flora.
Ano: 2004 URL: http://archimer.ifremer.fr/doc/2004/publication-579.pdf
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