Table 2.

PCR Primers, PCR targets, PCR annealing temperatures, and amplicon lengths (including primer sequences) for candidate nitrite reductase genes.

Primer Name and pair	Primer Sequence 5′→3′	Annealing temperature	Primer target Application	amplicon length
2967 EXP F (pair 1)	CAACAAATGTGAAATAAAGAG	53°C	BOGUAY_2967 and flanking regions	2483 bp
2967 EXP R (pair 1)	TCTAAAAATAAATGTGGTCTG	53°C	BOGUAY_2967 and flanking regions	2483 bp
2386 EXP F (pair 2)	TCCACCTTAACTAACTCACT	57°C	BOGUAY_2386 and flanking regions	1878 bp
2386 EXP R (pair 2)	GAATAAATATCAGAGCCGCT	57°C	BOGUAY_2386 and flanking regions	1878 bp
0691 EXP F (pair 3)	AAAAATGGTTGGCAGGTGGG	57°C	BOGUAY_0691 and flanking regions	1797 bp
0691 EXP R (pair 3)	ACTTTGCACCCCACATTACC	57°C	BOGUAY_0691 and flanking regions	1797 bp
2967F (pair 4)	ATGAGATATATTAGTCATTTCC	52°C	BOGUAY_2967	2016 bp
2967R (pair 4)	TTAATAAATGTCATGCGTCG	52°C	BOGUAY_2967	2016 bp
2386F (pair 5)	ATGATAAAAAAACAGCTCGC	52°C	BOGUAY_2386	1748 bp
2386R (pair 5)	TTTTTCACCTCCAATATGACAG	52°C	BOGUAY_2386	1748 bp
0691F (pair 6)	ATGATTCGTAAATTGTGGGC	54°C	BOGUAY_0691	1536 bp
0691R (pair 6)	TCTATCGGTTTGCTCACCATA	54°C	BOGUAY_0691	1536 bp
2967 FRE (pair 7)	CCCGTTCCATGGATGAGATATATTAGTCATTTCC	59°C	NcoI restriction site on BOGUAY_2967	2046 bp
2967 RRE (pair 7)	AAAGGGCCATGGCTCGAGATAAATGTCATGCGTCG	59°C	XhoI restriction site on BOGUAY_2967	2046 bp
2386 FRE (pair 8)	CCCGGGCCATGGATGATAAAAAAACAGCTCGC	63°C	NcoI restriction site on BOGUAY_2386	1778 bp
2386 RRE (pair 8)	AATAGTCCATGGCTCGAGTTTTTCACCTCCAATATGACAG	63°C	XhoI restriction site on BOGUAY_2386	1778 bp
0691 FRE (pair 9)	AAGCTTATGATTCGTAAATTGTGGGC	63°C	HindIII restriction site on BOGUAY_0691	1548 bp
0691 RRE (pair 9)	CTCGAGTCTATCGGTTTGCTCACCATA	63°C	XhoI restriction site on BOGUAY_0691	1548 bp
T7prom	TAATACGACTCACTATAGGG	60°C	pET22b Promoter region