Full genome seq of MBLV
The virus was isolated using the mouse inoculation test and viral RNA was then extracted using TRIzol (Invitrogen) (5). First-strand cDNA synthesis was performed using random hexamer primers (Integrated DNA Technologies) and the SuperScript III (Thermo Fisher Scientific) synthesis system. The full genome was amplified using four different primer sets (Table 1). Amplification reactions and cycling conditions were prepared using the Phusion High-fidelity DNA Polymerase system (New England Biolabs). Briefly, 1-unit Phusion DNA Polymerase, 0.5 M of each primer, 200 M dNTPs, 1X HF buffer and 3% DMSO were used. Next generation sequencing (NGS) was performed for each purified amplicon using the Nextera XT library prep kit (Illumina) on an Illumina MiSeq sequencing platform (Illumina) with paired end read lengths of 2 x 300 bp with 500 X coverage