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Drug Target Insights 2025 | DOI: 10.33393/d .2025.3592 | Smri  et al 

Supplementary Table 1: Primers used for aminoglycoside modifying enzymes (AMEs) genes in 
the study  

 

 

Forward/ 
Reverse 

Primer Sequences Amplic
on size 
(bp) 

References 

AAC(6’)-Ib F 5’TTGCGATGATGTATGACTGGCTA3’ 482bp Djagbare P 
et al., 2023 
(20) 

 R 5’CTCGAATCGCTGGCTTGAAA3’ 

AAC(3)-IIc F 5’ATATGCGAGAGTCGTAGCCGA3’ 877 bp Djagbare P 
et al., 2023 
(20) 

 R 5’GAGCCGCTCTGACCGGCAGG3’ 

APH(2”)-Ib F 5’CAAGCACGCTGAGCTATGTGAGCAG3’ 867bp Vakulenko et 
al., 2003 
(21) 

 R 5’GCCCGTAGGAATTCGAAAACGCCCTT3’ 

ANT(3”)-I F 5’TGACCCGCTGCATACGGTGAG3’ 284bp Wangkheim
ayum J et 
al., 2020 
(22) 

 R 5’CGGTGTGTTCTATTCCTTTTA3’ 

APH(3’)-Ia F 5’AATCGACTTGATCGAGGA3’ 461bp Wangkheim
ayumJ et al., 
2020 (22) 

 R 5’CAAGCCGTTATTGATACGTCA3’ 



Drug Target Insights 2025 | DOI: 10.33393/d .2025.3592 | Smri  et al 

Supplementary Table 2: Thermal cycling condi ons for standardiza on of PCR 

Ini al 
Denatura on 

Denatura on Annealing 
(Ta) 

Extension Final 
Extension 

Hold 

95°C for 5mins 95°C for1min AAC(6’)-Ib 

55°C for 
45sec 

72°C for 1min 72°C for 
5mins 

4°C ∞ 

95°C for 5mins 95°C for 1min AAC(3)-IIc 

57°C for 
45sec 

72°C for 1min 72°C for 
5mins 

4°C ∞ 

95°C for 5mins 95°C for 1min APH(2”)-Ib 

57°C for 
45sec 

72°C for 1min 72°C for 
5mins 

4°C ∞ 

95°C for 5mins 95°C for 1min ANT(3”)-I 

55°C for 
45sec 

72°C for 1min 72°C for 
5mins 

4°C ∞ 

95°C for 5mins 95°C for 1min APH(3’)-I 

55°C for 
45sec 

72°C for 1min 72°C for 
5mins 

4°C ∞