Sequencing of Dobrava-Belgrade orthohantavirus
The amplification reaction of the three segments (S, M and L) of DOBV sample using four pairs (one for S, one for M and two for L) of DOBV specific primers was performed in a two-step RT-PCR reaction using SuperScript IV VILO Master Mix with ezDNase Enzyme (Invitrogen) and Platinum SuperFi II Green PCR Master Mix (Invitrogen). The two samples were sequenced using next generation sequencing implemented in Illumina MiSeq with Reagent Nano Kit v2 (500 cycles).