Genomic Sequencing of Bank Vole-Derived BvHV Strains

For the determination of the genomic sequence of original or bank vole passaged BvHV strains (FA07, p1-KS13/920, p1-KS13/914), RNA was extracted from 500 µL BvHV-infected bank vole sera or liver tissue using TRIzol^® Reagent (Invitrogen, Darmstadt, Germany) and RNeasy Mini spin columns (Qiagen, Hilden, Germany) as previously described [36 (link)]. cDNA synthesis and library preparation were performed as previously described [37 (link)]. Sequencing was performed with an Illumina MiSeq instrument (Illumina, San Diego, CA, USA). Raw sequence data were analyzed and mapped using the Genome Sequencer software suite (v. 2.8; Roche, Mannheim, Germany) and Geneious software suite (v. 6.1.6; Biomatters, Auckland, New Zealand). Classification of the BvHV isolates was performed by phylogenetic analyses of complete genome sequences including reference strains MN242371.1 and KC411777.1 [25 (link)] using maximum-likelihood methods in MEGA5 [38 (link)] (Table S6).