Genes | Primer | Base pair | PCR reaction mixture | Temperature profile and Reference |
---|---|---|---|---|
Bla NDM−1 |
F = GGT TTG GCG ATC TGG TTT TC R=(CGG AAT GGC TCA TCA CGA TC |
621 bp | 8.5 µl of 2X master mix, 0.5 µl of 10 picomolar primer (forward and reverse), 12.5 µl of nuclease free water and 3 µl of extracted DNA template | initial denaturation at 94 °C for 5 min, followed by 36 cycles of 95 °C for 30 s, 52 °C for 40 s and 72 °C for 50 s with final extension at 72 °C for 5 min [18]. |
Mcr-1 |
F = CGGTCAGTCCGTTTGTTC R = CTTGGTCGGTCTGTAGGG |
309 bp | 21 µl 1X master mix, 0.5 µl of 10 pmolar primer (forward and reverse) and 3 µl of extracted plasmid DNA |
Initial heating at 95 °C for 15 min, then 35 cycles of 94 °C for 10 s, 57 °C for 90 s and 72 °C for 90 s and final extension at 72 °C for 10 min. [19] |
MexB |
F = TGTCGAAGTTTTTCATTGATAG R = AAGGTCAC GGTGATGGT |
280 bp | 21 µl of 1X Qiagen master mix, 0.5 µl of 10 pmole primers (forward and reverse) and 3 µl of extracted DNA template. | Initial heating at 94 °C for 3 min, then 32 cycles of 94 °C for 30 s, 55 °C for 45 s and 72 °C for 1 min and final extension at 72 °C for 7 min [20]. |