2.5 PCR amplification of LSDV P32 and F gene:
For amplification of LSDV genes, OIE recommended primers were employed (Table -1) (OIE, 2018; Sudhakar et al., 2020). For PCR, 12.5µl of 2X GoTaq Green master mix (GoTaq® DNA polymerase in 2X Green GoTaq® reaction buffer of pH 8.5, 400µM of each dNTPs and 3mM MgCl2) (Promega, USA), 1µl of each forward and reverse primer (10pmol), 150ng of DNA with final volume of 25µl was prepared using nuclease-free water. The reaction was carried out in gradient thermocycler (Eppendorf, Germany) with the conditions mentioned in Table-1. The PCR products were electrophoresed in 1.5% agarose gel and visualized under UV transilluminator. DNA from goat pox vaccine used as positive control in PCR reaction.