Gene | Method | Fragment | Primer name | Sequence (5′–3′) | PCR condition | |||
---|---|---|---|---|---|---|---|---|
Annealing Temperature (°C) | MgCl2 (mM) | No. of PCR cycle | Estimated PCR product Size (bp) | |||||
Pvkelch12 | PCR | Nest 1 | K12_F1-F | CCATACGTAAACGCTGCAAAT | 58 | 2 | 25 | 2139 |
K12_c.2216R | CTCCCCATCTGTTCCATGTC | |||||||
F1 | K12_F1-F | CCATACGTAAACGCTGCAAAT | 58 | 2 | 30 | 980 | ||
K12_F1-R* | TTCTTAATTTGTTTATACCCGTTTGA | |||||||
F2 | K12_F2-F | CAAGCTTTTTAAAGACAAAAAGGAA | 58 | 2 | 30 | 728 | ||
K12_F2-R* | CAGTTTCGAAAAGGGCTTTATAATC | |||||||
F3 | K12_c.1304F* | TGGTTTCGATGGGGTAGAGT | 58 | 2 | 30 | 912 | ||
K12_c.2216R | CTCCCCATCTGTTCCATGTC | |||||||
Pvpm4 | PCR | Nest 1 | PV_PM4F1_F | TCAAAAGGAGTACGAAGCATACAA | 62 | 2 | 30 | 1703 |
PV_PM4F2_R | TGTTCTAATTACAGCACCAACACA | |||||||
F1 | PV_PM4F1_F* | TCAAAAGGAGTACGAAGCATACAA | 62 | 2 | 30 | 807 | ||
PV_PM4F1_R* | ATGGGTTCTAAATCATCAGTGTCA | |||||||
F2 | PV_PM4F2_F* | GATGCAGCATTAAAAATCTGTACG | 62 | 2 | 30 | 1086 | ||
PV_PM4F2_R | TGTTCTAATTACAGCACCAACACA | |||||||
Pvpm4 | Vector-based primer | Pvpm4 | CPvPM4-F* CPvPM4-R* | AAAATCGAGAGACCCTATGACAAG ATGGGTTCTAAATCATCAGTGTCA | 62 | 2 | 35 | 461 |
Pv β-tubulin | CPvtubulin-F* | AAATTAGGGAAGAATACCCAGACC | 62 | 2 | 35 | 451 | ||
CPvtubulin-R* | TCACTTGCACACATCATATTCTTG | |||||||
Pvpm4 | Real-time PCR | Pvpm4 | Pvpm4_F(CNV) Pvpm4_R(CNV) PvPm4_Probe(CNV) | ACCACCAAAAGTTTATGCTCATC CGTAAGACTTAGACTTGCTAGAGTCG 6FAM-CAAAAAATGCAATAGCAGCGGA-TAMRA | 58 | – | 50 | 133 |
Pv β-tubulin | PvTubulin_F(CNV) PvTubulin_R2(CNV) PvTubulin_Probe(CNV) | GATAATGAGGCCTTGTATGATATTT AGGAAATCTTAACGAACATGTAACT HEX-TGGTTTCTGCTGCCATGTCAGG-TAMRA | 58 | – | 50 | 123 |