Table 1.
Primer pairs used to amplify and sequence Pfcrt and Pfmdr1
| Gene | Primer | Primer sequence | PCR Cycling conditions |
|---|---|---|---|
|
P. falciparum Genus P. falciparum |
rPLU1 | TCAAAGATTAAGCCATGCAAGTGA |
25 cycles of 94 °C/1 min, 58 °C/1 min, and 72 °C/2 min 30 cycles of 94 °C/1 min, 58 °C/1 min, and 72 °C/2 min |
|
rPLU5 rFAL1 rFAL2 |
CCTGTTGTTGCCTTAAACTTC TTAAACTGGTTTGGGAAAACCAAATATATT ACACAATGAACTCAATCATGACTACCCGTC |
||
|
Pfcrt SNPs Codon 72 and 76 |
OF P1 | 5′-GACGAGCGTTATAGAGAATTA-3′ | 35 cycles of 94 °C/30 s; 56 °C/30 s; and 62 °C/1 min |
| OR P2 | 5′-CCAGTAGTTCTTGTAAGACC-3′ | ||
| NF P3 | 5′-GGCTCACGTTTAGGTGGA-3′ | 30 cycles of 94 °C/30 s; 56 °C/30 s; and 65 °C/1 min | |
| NR P4 | 5′-TGAATTTCCCTTTTTATTTCCAAA3′ | ||
|
Pfmdr1 SNPs Codon 86 and 184 |
OF P5 | 5′-AGGTTGAAAAAGAGTTGAAC-3′ | 30 cycles of 94 °C/30 s; 55 °C/30 s; and 65 °C/1 min |
| OR P6 | 5′-ATGACACCACAAACATAAAT-3′ | ||
| NF P7 | 5′-ACAAAAAGAGTACCGCTGAAT-3′ | 30 cycles of 94 °C/30 s; 60 °C/30 s; and 65 °C/1 min | |
| NR P8 | 5′AAACGCAAGTAATACATAAAGTC3′ | ||
|
Pfmdr1 SNPs Codon 1034 1042 & 1246 |
OF P9 | 5′-GTGTATTTGCTGTAAGAGCT-3′ | 34 cycles of 94 °C/30 s; 55 °C/60 s and 72 °C/1.5 min |
| OR P10 | 5′GACATATTAAATAACATGGGTTC | ||
| NF P11 | 5′CAGATGATGAAATGTTTAAAGATC | 29 cycles of 94 °C/30 s; 60 °C/30 s; and 65 °C/1 min | |
| NR P12 | 5′-TAAATAACATGGGTTCTTGACT |