Bacteriaweremaintained at−80 °C and resuscitated inM17 broth (Difco) containing 2% lactose or glucose (w/v) for S. thermophilus, S.salivarius, and L. garvieae and in MRS broth (Difco) for all Lactobacillus species. The cultures were incubated at 37 °C and 30 °C for thermophilic and mesophilic species, respectively. The identification of isolates at the species level was performed by means of 16S rDNA gene sequencing. The sequences of the primers used for the amplification of the 16S rDNA gene were P0 and P6, and the PCR protocol described by the authors was used (Di Cello and Fani, 1996). S. thermophilus and S. salivarius strains were identified by secY gene sequencing (Pombertet al., 2009). The PCR products were purified using an UltraClean DNA Purification Kit (MO BIO). Finally, sequence comparison was per-
formed with Genetics Computer Group sequence analysis software
using BLAST.
Bacteriaweremaintained at−80 °C and resuscitated inM17 broth (Difco) containing 2% lactose or glucose (w/v) for S. thermophilus, S.salivarius, and L. garvieae and in MRS broth (Difco) for all Lactobacillus species. The cultures were incubated at 37 °C and 30 °C for thermophilic and mesophilic species, respectively. The identification of isolates at the species level was performed by means of 16S rDNA gene sequencing. The sequences of the primers used for the amplification of the 16S rDNA gene were P0 and P6, and the PCR protocol described by the authors was used (Di Cello and Fani, 1996). S. thermophilus and S. salivarius strains were identified by secY gene sequencing (Pombertet al., 2009). The PCR products were purified using an UltraClean DNA Purification Kit (MO BIO). Finally, sequence comparison was per-formed with Genetics Computer Group sequence analysis softwareusing BLAST.
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