|
|
|
|
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 |
|
|
|
|
|