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.