Selected article for: "GenBank database and rrna gene"
Title: 2016 ACVIM Forum Research Abstract Program
  • Document date: 2016_5_31
    • ID: 2y1y8jpx_789
    Snippet: Composite milk samples were aseptically collected from all 940 lactating goats and aerobic bacteriological culture was performed. A culture yielding 100 CFU/mL CNS was considered positive. Isolates from milk samples yielding a single colony morphology identified as CNS were used to prepare lysates for PCR amplification. Species identification was based on PCR amplification and sequencing of either the rpoB, tuf gene, or 16S rRNA. Sequences were c.....
    Document: Composite milk samples were aseptically collected from all 940 lactating goats and aerobic bacteriological culture was performed. A culture yielding 100 CFU/mL CNS was considered positive. Isolates from milk samples yielding a single colony morphology identified as CNS were used to prepare lysates for PCR amplification. Species identification was based on PCR amplification and sequencing of either the rpoB, tuf gene, or 16S rRNA. Sequences were compared with the GenBank database using the NCBI Nucleotide-BLAST algorithm. For rpoB, species identification was assigned with ≥ 97% identity and ≥ 5% separation between different species. For tuf, species identification was assigned with ≥ 98% identity and > 0.8% separation between different species. For 16S, species identification was assigned with ≥ 99% identity and > 0.8% separation between different species. PCR amplification and sequencing of rpoB, tuf and 16S were performed sequentially until identification was assigned. For instance, tuf was performed on isolates for which rpoB either yielded unsuccessful amplification, or criterion for identification with sequence analysis were not met, followed by 16S if tuf amplification or sequence analysis failed.

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