Genes names | Primers names | Primers sequences | amplicon size (bp) | PCR cycling conditions |
---|---|---|---|---|
Pfdhfr | ||||
PCR1 | dhfr_M1 | 5′-TTTATGATGGAACAAGTCTGC-3′ | 650 | 93°C for 5min/(94°C for 30s- 54°C for 60s- 65°C for 60s) × 41cycles/ 65° for 5min/ 15°C for 5min |
Nested | dhfr_M7 | 5′-CTAGTATATACATCGCTAACA-3′ |  |  |
 | dhfr-M9 | 5′-CTGGAAAAAATACATCACATTCATATG-3′ | 594 | 95°C for 5min/(93°C for 30s- 56°C for 30s- 68°C for 75s) × 30 cycles/ 75°C for 5min |
Sequencing | dhfr-M3 | 5′-TGATGGAACAAGTCTGCGACGTT-3′ |  |  |
 | dhfr-M9 | 5′-CTGGAAAAAATACATCACATTCATATG-3′ |  | 96°C for 1min/(96°C for 30s- 50°C for 30s- 60°C for 4min) × 26 cycles/4°C hold until ready to purify |
 | dhfr-M3 | 5′-TGATGGAACAAGTCTGCGACGTT-3′ |  |  |
Pfdhps | ||||
PCR1 | dhps-N1 | 5′-GATTCTTTTTCAGATGGAGG-3′ | 770 | 94°C for 3min/(94°C for 30s- 55°C for 30s- 65°C for 60s) × 30cycles / 65°C for 5min/ 15°C for 5min. |
 | dhps-N2 | 5′-TTCCTCATGTAATTCATCTGA-3′ | 711 |  |
Nested | dhps-R2 | 5′-AACCTAAACGTGCTGTTCAA-3′ |  | 94°C for 5min/(94°C for 30s- 60°C for 30s- 65°C for 1min) × 30cycles /65°C for 5min/15°C for 5min. |
 | dhps-R/ | 5′-AATTGTGTGATTTGTCCACAA-3′ |  |  |
 | dhps-R2 | 5′-AACCTAAACGTGCTGTTCAA-3′ |  | 96°C for 1min/(96°C for 30s-50°C-60°C for 4min) × 26 cycles/ 4°C hold until ready to purify. |
Sequencing | dhps-R/ | 5′-AATTGTGTGATTTGTCCACAA-3′ |  |  |
Pfmdr1 | ||||
PCR1 | fn1/1 | 5′-ACAAAAAGAGTACCGCTGAAT-3′ | 578 | 94°C for 3min/(94°C for 30s- 55°C for 30s- 65°C for 1min) × 30cycles/ 65°C for 5min/ 15°C for 5min. |
 | rev/c1 | 5′-AAACGCAAGTAATACATAAAGTC-3′ | 534 |  |
Nested | mdr2/1 | 5′-ACAAAAAGAGTACCGCTGAAT-3′ |  | 94°C for 3min/(94°C for 30s- 60°C for 30s- 65°C for 1min) × 30cycles/ 65°C for 5min/ 15°C for 5min. |
 | newrev1 | 5′-AAACGCAAGTAATACATAAAGTC-3′ |  |  |
Sequencing | mdrfr1f-seq | 5′-GTCGAATTATTTAGAAAAAT-3′ |  | 96°C for 1min/(96°C for 30s- 50°C -60°C for 4min) × 26cycles/4°C hold until ready to purify. |
 | mdrfr1r-seq | 5′-GCAAGTAATACATAAAGT-3′ |  |  |
Pfcrt | ||||
 | Crtd1 | 5′-TGTGCTCATGTGTTTAAACTT 5′- | 166 | 94°C for 3min/(94°C for 30s- 55°C for 30s- 65°C for 1min) × 30 cycles/ 65°C for 5min/15°C for 5min. |
 | Crtd2 | CAAAACTATAGTTACCAATTTTG |  |  |