Viral detection by high-throughput sequencing

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1236; p. 125
• Main Authors: Motooka, Daisuke, Nakamura, Shota, Hagiwara, Katsuro, Nakaya, Takaaki
• Format: Journal Article
• Language: English
• Published: United States 01.01.2015
• Subjects: Animals; Bacteriophages - genetics; Bacteriophages - isolation & purification; Cattle; DNA - isolation & purification; DNA, Complementary; Feces - virology; Female; Gene Library; Genome, Viral; High-Throughput Nucleotide Sequencing - instrumentation; High-Throughput Nucleotide Sequencing - methods; Insect Viruses - genetics; Insect Viruses - isolation & purification; Reverse Transcriptase Polymerase Chain Reaction - methods; RNA - isolation & purification; Viruses - genetics; Viruses - isolation & purification

We applied a high-throughput sequencing platform, Ion PGM, for viral detection in fecal samples from adult cows collected in Hokkaido, Japan. Random RT-PCR was performed to amplify RNA extracted from 0.25 ml of fecal specimens (N = 8), and more than 5 μg of cDNA was synthesized. Unbiased high-throug...