Table 5.
List of primers used with the real-time RT-PCR HRM assay and the end-point multiplex RT-PCR protocol
| Primer pair | Sequence (5'-3') | Target region | Amplicon size (bp) |
|---|---|---|---|
| LR3.HRM1.F |
TAGACGTTAAAGATGTGAAGCG |
GLRaV-3 ORF1a |
167 |
| LR3.HRM1.R |
TCGTACACATCCACCATA |
|
|
| LR3.HRM2.F |
GTCCTAGATTCGGATTTTGTCG |
GLRaV-3 ORF1a |
231 |
| LR3.HRM2.R |
GAATACTCTTCGCCCTATC |
|
|
| LR3.HRM3.F |
CTGGTTGCTTTCGAGGTATATGAG |
GLRaV-3 ORF1b |
295 |
| LR3.HRM3.R |
CACTTCAAGGTGTTGCGCTT |
|
|
| LR3.HRM4.F |
TAATCGGAGGTTTAGGTTCC |
GLRaV-3 ORF4 |
226 |
| LR3.HRM4.R |
GTCGGTTCGTTAACAACAC |
|
|
| LR3.HRM5.F |
TGTGTAAGAAGGTTATGGG |
GLRaV-3 ORF6 |
224 |
| LR3.HRM5.R |
TACTGCCTTACCGGGTTTTC |
|
|
| LR3.HRM6.F |
GTCACCAGGTGTTCCAAACC |
GLRaV-3 ORF1a |
305 |
| LR3.HRM6.R |
AACGCCCTGTATGTCCTCTC |
|
|
| LR3_Universal_F |
TAAATGCTCTAGTAGGATTC |
GLRaV-3 5'UTR |
|
| 621_430R |
TAACCCAACACGACGATGAG |
GLRaV-3 5'UTR |
429a |
| 623_564R |
CTCACGCTAAACACACCAAG |
GLRaV-3 5'UTR |
563a |
| PL20_315R |
GTTTGTAACAAAGAAACACG |
GLRaV-3 5'UTR |
314a |
| GH11_180R |
CCAAAACGAAGACGAAAAGAAGAG |
GLRaV-3 5'UTR |
179a |
| LR_ORF1aR |
CGTCCGCTTCACCCCTTTGG |
GLRaV-3 ORF1a |
|
| Vv_Actin_F [32] |
CTTGCATCCCTCAGCACCTT |
V. vinifera predicted actin-7 |
82 |
| Vv_Actin_R [32] | TCCTGTGGACAATGGATGGA |
aAmplicon size if use together with LR_Universal_F.