No Gene primers details (5’ to 3’) Size (bp) Cyclic conditions Cyclic conditions Cyclic conditions Reference
Denaturation Annealing Extension
1 RPO30-F CTC TGT TCC AAA CTA AAT CAT 1385 95oC for 1 min 49oC for 1 min 72oC for 1 min OIE, 2018; Sudhakar et al., 2020
2 RPO30-R TTT TTG TAT TAC CAA TTT CTG
3 P32-F TCC GAG CTC TTT CCT GAT TTT TCT TAC TAT 192 95oC for 1 min 49oC for 1 min 72oC for 1 min OIE, 2018; Ireland &Binepal, 1998
4 P32-R TAT GGT ACC TAA ATT ATA TAC GTA AAT AAC
5 F gene (LSDV117)-F ACT AGT GGA TCC ATG GAC AGA GCT TTA TCA 470 95oC for 1 min 49oC for 1 min 72oC for 1 min
6 F gene (LSDV117) R GCT GCA GGA ATT CTC ATA GTG TTG TAC TTC G
TaqMan Probe and primers
TaqMan Probe and primers
UDG Incubation
Polymerase activation
Denaturation
Annealing/Extension
OIE, 2018; Pestova et al., 2018
7 LSD f2 TAG GAA AAT GGA TGT ACC ACA AAT ACA G 50oC 2 min 95oC for 10 min 95oC for 15 sec 60oC for 1 min
8 LSD r33 TTG TTA CAA CTC AAA TCG TTA GGT G
9 TaqMan probe FAM - ACC ACC TAA TGA TAG TGT TTA TGA TTT AC- 31 BHQ1