gene | Primer sequence (5'-3') | Length of amplicon | Thermal profile for PCR |
amoA AOB amoA gene | amoA-lF: GGG GTT TCT ACT GGT GGT amoA-2R: CCC CTC KGS AAA GCC TTC TTC | 491 bp | 95˚C, 3.0 min; 35× (95˚C, 0 s; 55˚C, 20 s; 72˚C, 20 s, 83˚C, 20 s with plate read); Melt curve 65.0˚C to 95.0˚C, increment 0.5˚C 0:05+ plate read |
amoA AOA amoA gene | Arch-amoAF; TAATGG TCTGGC TTA GAC G Arch-amoAR: CGG CCA TCC ATC TGT ATG T | 635 bp | 95˚C, 3.0 min; 39× (95˚C, 10 s; 55˚C, 20 s; 72˚C, 20 s with plate read); Melt curve 65.0˚C to 95.0˚C, increment 0.5˚C, 0:05+ plate read |