Identification of papillomaviruses in bats
Of each of the fecal samples, 5 g was homogenized in SM buffer and the homogenate was centrifuged at 6000 g for 10 min. The supernatant was sequentially filtered through 0.45 m and 0.2 m syringe filters and viral particles in the filtrate were precipitated with 15% (w/v) PEG-8000 with overnight incubation at 4 C followed by centrifugation at 10,000 g. The pellet was resuspended in 500l of SM Buffer and 200L of this was used for viral DNA extraction using the High Pure Viral Nucleic Acid Kit (Roche Diagnostics, USA). The total DNA was amplified using rolling circle amplification (RCA) with TempliPhi 2000 kit (GE Healthcare, USA) and the RCA products used to prepare Illumina sequencing libraries and sequenced on an Illumina NovaSeq 6000 sequencer.
SUB9384548
UoAChi13