Gene | Amplicons | Fw primer | Rv primer | Amplification conditions |
---|---|---|---|---|
Dao (PR1) | + 3/+ 365 | TagTTagagaagtTaggYtgYtYaYta | agattggtgaRRRaaaaaaggagaga | Denature at 95 °C for 2 min; 35 cycles of denaturing at 95 °C for 30 s, annealing at 52 °C for 40 s, and extension at 72 °C for 50 s. Final elongation at 72 °C for 6 m |
Dao (PR2) | − 234/+ 137 | gaaYagYagTgagYtagYtgg | caccaRccaRRaatRaaacacaa | Denature at 95 °C for 2 min; 38 cycles of denaturing at 95 °C for 30 s, annealing at 54 °C for 40 s, and extension at 72 °C for 50 s. Final elongation at 72 °C for 6 m |
Srr | + 27/+ 377 | GTaTtgggagTaaaagTattTag | tttaaactccacaatccaAAcct | Denature at 95 °C for 2 min; 35 cycles of denaturing at 95 °C for 30 s, annealing at 57 °C for 40 s, and extension at 72 °C for 50 s. Final elongation at 72 °C for 6 m |
Ddo | − 63/− 468 | gtgtgtttTtgaggaggtgaTaTtTa | aActtaccctccattAAtccatAcc | Denature at 95 °C for 2 min; 36 cycles of denaturing at 95 °C for 30 s, annealing at 52 °C for 40 s, and extension at 72 °C for 50 s. Final elongation at 72 °C for 6 m |
M13mp18 | 5946/6294 | Ggtgaagggtaattagttgttgtt | ccaataccaaacttacatacct | Denature at 95 °C for 2 min; 33 cycles of denaturing at 95 °C for 30 s, annealing at 57 °C for 40 s, and extension at 72 °C for 50 s. Final elongation at 72 °C for 6 m |