Gene Primers* | Squencen(5′-3′) | PCR cycling conditions** |
---|---|---|
pvcsp(N1) | F:5′-ATGTAGATCTGTCCAAGGCCATAAA-3′ | 95°C 3 min/[94°C 30 s, 58°C 30 s, 72°C 1.5 min] × 30 cycles, 72°C 5 min |
R:5′-TAATTGAATAATGCTAGGACTAACAATATG-3′ | ||
pvcsp(N2) | F:5′-CCAGATGACGAGGAAGGAGATGC-3′ | 95°C 3 min/[94°C 30 s, 58°C 30 s, 72°C 1 min] × 35 cycles, 72°C 5 min |
R:5′-TCTTTCACAGACTTTTCATTTGGG-3′ | ||
pvmsp-1(N1) | F:5′-GAGCCCTACTACTTGATGGTCC-3′ | 95°C 3 min/[94°C 30 s, 58°C 30 s, 72°C1min] × 35 cycles, 72°C 5 min |
R:5′-CCTTCTGGTACAGCTCAATG-3′ | ||
pvmsp-3α(N1) | F:5′-CAGCAGACACCATTTAAGG-3′ | 95°C 3 min/[94°C 30 s, 54°C 30 s, 68°C 2.5 min] × 30 cycles, 68°C 5 min |
R:5′-CCGTTTGTTGATTAGTTGC-3′ | ||
pvmsp-3α(N2) | F:5′-GACCAGTGTGATACCATTAACC-3′ | 95°C 3 min/[94°C 30 s, 55°C 30 s, 68°C 2.5 min] × 40 cycles, 68°C 5 min |
R:5′-ATACTGGTTCTTCGTCTTCAGG-3′ | ||
pvmsp-3β(N1) | F:GTATTCTTCGCAACACTC | 95°C 3 min/[94°C 30 s, 54°C 30 s, 68°C 2.5 min] × 30 cycles, 68°C 5 min |
R:CTTCTGATGTTATTTCCAG | ||
pvmsp-3β(N2) | F:CGAGGGGCGAAATTGTAAACC | 95°C 3 min/[94°C 30 s, 55°C 30 s, 68°C 2.5 min] × 40 cycles, 68°C 5 min |
R:GCTGCTTCTTTTGCAAAGG |