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