title: Complete genome sequences of novel papillomaviruses in captive black-and-white ruffed lemurs (Varecia variegata )
design: Saliva samples were collected by allowing the lemurs to chew on a SalivaBio Childrens Swab (Salivametrics, USA). The saturated swabs were then placed into a SalivaBio Swab Storage Tube (Salivametrics, USA) and centrifuged to collect the saliva at the bottom of the storage tube. Saliva samples were stored at -80 until  viral DNA extraction. To the saliva sample, SM buffer (0.1M NaCl, 50mM Tris-HCl [pH 7.4]) was added in order to obtain 400 l of solution per sample. 200 l of solution per sample was run through the High Pure Viral Nucleic Acid Kit (Roche Diagnostics, USA) to perform viral DNA extraction. 1l of the extracted viral DNA was amplified using the Illustra TempliPhi rolling circle amplification kit (GE Healthcare, USA) to target circular viruses. The viral DNA and rolling amplified viral DNA were combined. Libraries were generated using the Illumina DNA Prep kit which utilizes tagmentation to fragment the DNA. Reactions were performed at a reduced 1/8 volume reaction on the SPT Labtech HV Mosquito and Dragonfly Discovery liquid handlers. Libraries were sequenced using the Illumina NovaSeq 6000 on an SP flowcell (2x150bp).