2.2 Viral RNA/DNA extractions
Viral RNA (FMDV, VSV, BTV and PPRV) was extracted from 140 μl of CC or
CS using the RNeasy Mini Kit (Qiagen, Valencia, CA) according to the
manufacturer’s instructions. Viral DNA (CaPV, PaPV and BHV) was
extracted from 100 μl of CC or CS using the DNeasy Blood and Tissue Kit
(Qiagen) according to the manufacturer’s instructions. Tissue and skin
samples were processed to 10% homogenates in Dulbecco’s Modified Eagle
medium (DMEM, Thermo Fisher Scientific) as described previously (Das et
al. 2017a). To minimize template degradation, extracted viral DNA/RNA
were appropriately diluted in nuclease-free water and aliquoted into a
smaller volume for one time use only and stored at -70 ºC.