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Comparison of three methods of DNA extraction from cold-smoked salmon and impact of physical treatments ArchiMer
Giacomazzi, Sophie; Leroi, Francoise; Joffraud, Jean-jacques.
Aims: To compare three bacterial DNA extraction procedures on cold-smoked salmon (CSS) and assess the impact on their efficiency of two physical treatments of the food matrix, ionizing irradiation and freezing. Methods and Results: As molecular methods for bacterial detection have become an important analytical tool, we compared bacterial DNA extraction procedures on CSS. Working with frozen and irradiated CSS, we obtained negative responses from samples known to be highly contaminated. Thus, we decided to study the impact of these two physical treatments on bacterial DNA extraction procedures. The efficiency of bacterial DNA extraction directly from the fish matrix suspension was measured by an rpoB PCR-based reaction. The results demonstrated that the...
Tipo: Text Palavras-chave: RpoB gene; PCR; Ionizing irradiation; Freezing; DNA extraction; Cold smoked salmon.
Ano: 2005 URL: http://archimer.ifremer.fr/doc/2005/publication-1106.pdf
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Identification of Leptospira serovars by RFLP of the RNA polymerase beta subunit gene (rpoB) BJM
Jung,Lenice Roteia Cardoso; Bomfim,Maria Rosa Quaresma; Kroon,Erna Geessien; Nunes,Álvaro Cantini.
Leptospires are usually classified by methods based on DNA-DNA hybridization and the conventional cross-agglutination absorption test, which uses polyclonal antibodies against lipopolysaccharides. In this study, the amplification of the rpoB gene, which encodes the beta-subunit of RNA polymerase, was used as an alternative tool to identify Leptospira. DNA extracts from sixty-eight serovars were obtained, and the hypervariable region located between 1990 and 2500-bp in the rpoB gene was amplified by polymerase chain reaction (PCR). The 600-bp amplicons of the rpoB gene were digested with the restriction endonucleases TaqI, Tru1I, Sau3AI and MslI, and the restriction fragments were separated by 6% polyacrylamide gel electrophoresis. Thirty-five fragment...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Leptospira; RpoB gene; RFLP; Serovar; DNA typing.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822015000200465
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