Table 1.

Quantitative PCR (qPCR) primers and conditions used for the amplifications of nosZI, nosZII, nirS, nifH, 16SrRNA gene.

Target	Forward primer	Reverse primer	Primer reference	Hold stage	PCR stage	Complete extension	Melt curve
nosZII	1153_nosZ8F- CTIGGICCIY TKCAYAC	1888_nosZ29R- GCIGAICARA AITCBGTRC	Jones et al., 2013	50°C, 95°C	95°C, 60–54°C, 72°C, 80°C	-	95°C, 60°C, 95°C, 60°C
				2 min, 10 min	15 s, 30 s, 30 s, 30 s	-	15 s, 1 min, 30 s, 15 s
					-44 cycles at 54°C annealing temperature
nifH	nifHF-AAAGGYGGWATCGG YAARTCCACCAC	nifHRb-TGSGCYTTGTCYTCR CGGATBGGCAT	Rösch and Bothe, 2005	96°C	96°C, 58.5°C, 68°C	72°C	96°C, 50°C, 96°C, 72°C
				10 min	3 s, 3 s, 30 s	5 min	15 s,1 min, 15 s, 5 min
					-40cycles
nosZI	nosZIF-CGCRACGGCAAS AAGGTSMSSGT	nosZIR-CAKRTGCAKSGC RTGGCAGAA	Henry et al., 2006	96°C	96°C, 58.5°C, 68°C	72°C	96°C, 50°C, 96°C
				10 min	3 s, 3 s, 30 s	5 min	15 s, 1 min, 15 s
					-40 cycles
nirS	cd3aF-GTSAACGTSAA GGARACSGG	R3cd-GASTTCGGRTGS GTCTTGA	Throback et al., 2004	95°C	95°C, 58.5°C, 72°C, 77°C	72 C	95°C, 60°C, 95°C
				10 min	10 s, 20 s, 20 s, 20 s -40 cycles	5 min	15 s, 1 min, 15 s
16S rRNA gene	16S UNIV-F-ACT CCT ACG GGA GGC AGC AG	16S UNIV-R-ATT ACC GCG GCT GCT GGC	Hartman et al., 2009	95°C	95°C, 65°C, 72°C	-	95°C, 60°C, 95°C, 72°C
				15 s	15 s, 20 s, 20 s -40 cycles	-	15 s, 1 min, 15 s, 5 min

Annealing temperatures are in bold.