Table 1 Pfmdr1 PCR primer sequences and reaction conditions used in Fragments 1, 3 and 4 amplification reactions
Gene fragment | Primer name | Â | Primer sequence | Codons | PCR cycling conditions |
|---|---|---|---|---|---|
Fragment 1 | Â | Â | Â | Â | Â |
Primary FR1 | FN1/1 | F | 5′- AGGTTGAAAAAGAGTTGAAC-3′ | 86, 184 | 94°C 3 min/[94°C 30 s-45°C 60 s 72°C 60 s] |
|  | REV/C1 | R | 5′- ATGACACCACAAACATAAAT-3′ |  | ×30 cycles |
Nested FR1 | MDR2/1 | F | 5′- ACAAAAAGAGTACCGCTGAAT -3′ |  | 72°C for 5 minutes/15°C 5 min |
|  | NEWREV1 | R | 5′-AAACGCAAGTAATACATAAAGTC-3′ |  |  |
Fragment 3 | Â | Â | Â | Â | Â |
Primary FR3 | MDRFR3N1 | F | 5′-GCATTTTATAATATGCATACTG-3′ | 1034, 1042 | 94°C 3 min/[94°C 30 s-55°C 60 s 65°C 40 s] |
|  | MDRFR3R1 | R | 5′-GGATTTCATAAAGTCATCAAC-3′ |  | ×30 cycles |
Nested FR3 | MDRFR3N2 | F | 5′-GGTTTAGAAGATTATTTCTGTA-3′ |  | 72°C 5 min/15°C 5 min |
|  | MDRFR3R1 | R | 5′-GGATTTCATAAAGTCATCAAC-3′ |  |  |
Fragment 4 | Â | Â | Â | Â | Â |
Primary FR4 | MDRFR4N1 | F | 5′- CAAACCAATCTGGATCTGCAGAAG -3′ | 1246 | 94°C 3 min/[94°C 30 s-55°C 60 s-65°C 40 s] |
|  | MDRFR4R1 | R | 5′-CAATGTTGCATCTTCTCTTCC -3′ |  | ×30 cycles |
Nested FR4 | MDRFR4N2 | F | 5′- GATCTGCAGAAGATTATACTG -3′ |  | 72°C 5 min/15°C 5 min |
|  | MDRFR4R1 | R | 5′- CAATGTTGCATCTTCTCTTCC -3′ |  |  |