Table 2 PCR condition and primer sets used for the screening of Vibrio virulence toxins (genes).
Virulence toxin (genes) | PCR primer sequence (5′–3′) | Amplicon size (bp) | PCR cycling condition | References |
|---|---|---|---|---|
tdhF | GGTCTAAATGGCTGACATC | 199 | Initial denaturation (93 °C for 5 min), denaturation (92 °C for 30 s), 35 cycles, annealing (55 °C for 60 s), extension (72 °C for 60 s), final extension (72 °C for 7 min) | |
tdhR | CCACTACCACTCTCATATGC | |||
trhF | CATTTCCGCTCTCATATGC | 250 | ||
trhR | GGCTCAAAATGGTTAAGCG | |||
vcgCP1 | AGCTGCCGATAGCGATCT | 278 | Initial denaturation (93 °C for 5 min), denaturation (94 °C for 40 s), 35 cycles, annealing (56 °C for 40 s), extension (72 °C for 60 s), final extension (72 °C for 7 min) | |
vcgP3 | CGCTTAGGATGATCGGTG | |||
vcgEP2 | CTCAATTGACAATGATCT | 278 | Initial denaturation (94 °C for 5 min), denaturation (94 °C for 40 s), 35 cycles, annealing (49 °C for 40 s), extension (72 °C for 60 s), final extension (72 °C for 7 min) | |
vcgP3 | CGCTTAGGATGATCGGTG | |||
vfh-F | GCGCGTCAGTGGTGGTGAAG | 800 | Initial denaturation (94 °C for 15 min), denaturation (94 °C for 40 s), 35 cycles, annealing (50–60 °C for 40 s), extension (72 °C for 60 s), final extension (72 °C for 7 min) | |
vfh-R | TCGGTCGAACCGCTCTCGCTT | |||
hupO-F | ATTACGCACAACGAGTCGAAC | 600 | Initial denaturation (93 °C for 15 min), denaturation (92 °C for 40 s) 35 cycles, annealing (50–62 °C for 60 s) extension (72 °C for 90 s) final extension (72 °C for 7 min) | |
hupO-R | ATTGAGATGGT AAACAGCGCC | |||
vfpA-F | TACAACGTCAAGTTAAAGGC | 1790 | ||
vfpA-R | GTAGGCGCTGTAGCCTTTCA | |||
stn-F | GGTGCAACATAATAAACAGTCAACAA | 375 | ||
stn-R | TAGTGGTATGCGTTGCCAGC |
