Selected article for: "direct sequencing and GenBank database"
Author: SOMA, Takehisa; MATSUBAYASHI, Makoto; SASAI, Kazumi
Title: Detection of kobuvirus RNA in Japanese domestic dogs
  • Document date: 2016_8_2
    • ID: 5b8i71jd_8
    Snippet: Furthermore, 12 samples positive for the above mentioned RT-PCR were also submitted to RT-PCR with a primer pair which was amplified at nucleotide positions 6,982-7,485 (3D gene) of CaKoV 12D049 (KF924623) (Forward; 5′-CCCTGGAACACCCAAGGCCGCT-3′, Reverse; 5′-TCTGGTTGCCATAGATGTGGTG-3′) [5] . These PCR amplicons (504-bp) and above amplicons (252-bp) were purified using ExoSAP-IT and were submitted to direct sequencing on both strands by the .....
    Document: Furthermore, 12 samples positive for the above mentioned RT-PCR were also submitted to RT-PCR with a primer pair which was amplified at nucleotide positions 6,982-7,485 (3D gene) of CaKoV 12D049 (KF924623) (Forward; 5′-CCCTGGAACACCCAAGGCCGCT-3′, Reverse; 5′-TCTGGTTGCCATAGATGTGGTG-3′) [5] . These PCR amplicons (504-bp) and above amplicons (252-bp) were purified using ExoSAP-IT and were submitted to direct sequencing on both strands by the dye-terminator cycle sequencing methodology (Bio Matrix Research, Chiba, Japan). The sequences (Nucleotide positions 6,982-7,619; 638-bp) were aligned with the reference strains in the GenBank Database using MEGA 6 software [27] . Phylogenetic trees were constructed employing the neighbor-joining method [23] , and their homologies were analyzed using the BLAST search.

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