Table 1.

Primers and PCR conditions used for tetracycline resistance gene amplification

Gene	Primers	Primer sequences 5′-3′ (Reference)	PCR cycles	Amplicon (bp)	Positive control
otr(A)	otr(A) (F) otr(A) (R)	GAACACGTACTGACCGAGAAG CAGAAGTAGTTGTGCGTCCG (37)	5 min/95°C; 35×(1 min/94°C, 30 s/60°C, 30 s/72°C); 5 min/72°C	778	Streptomyces rimosus subsp. rimosus DSMZ 40260 (ATTC 10970)
otr(B)	otr(B) (F) otr(B) (R)	CCGACATCTACGGGCGCAAGC GGTGATGACGGTCTGGGACAG (37)	5 min/95°C; 35×(1 min/94°C, 1 min/68°C, 1 min/72°C); 7 min/72°C	947	Streptomyces rimosus subsp. rimosus DSMZ 40260 (ATTC 10970)
tap	Tap1 Tap2	GTCGCGTTCCCGTGGCTGGT CGATACCGGGGCCGACGATG (22)	10 min/94°C; 35×(1 min/94°C, 30 s/68°C, 30 s/72°C); 3 min/72°C	400	Mycobacterium fortuitum TR-1242 (this study)
tet(K) and tet(L)	tetKL-FW tetKL-RV	TTACCTGATATTGCAA GACCAATGAATATAAT (this study)	5 min/95°C; 35×(30 s/94°C, 30 s/40°C, 30 s/72°C); 3 min/72°C	397	Staphylococcus haemolyticus CB-N (tet(K); this study) and Staphylococcus aureus pSTS9-like (tet(L); 1)
tet(M)	TetM-FW TetM-RV	ACAGAAAGCTTATTATATAAC TGGCGTGTCTATGATGTTCAC (6)	4 min/94°C; 35×(20 s/94°C, 30 s/52.3°C, 1 min/72°C); 7 min/68°C	171	Plasmid pAT101 that carries tet(M) gene from Streptococcus transposon Tn1545 (34)
tet(V)	tetV-FW tetV-RV	GCCTACGGTTTCATCCTGGC CGAGACCACCTTCGACAGCG (this study)	7 min/95°C; 35×(1 min/94°C, 15 s/65°C, 30 s/72°C); 5 min/ 72°C	351	Mycobacterium sp. Site2-2C (this study)