TABLE 1.

PCR primer sequences and reaction conditions used for the amplification of sequences containing P. falciparum pfcrt and pfmdr1 genes

Gene	Primer name (positions)	Primer sequence	Product size (bp)	Restriction endonuclease	PCR cycling conditions
Nest I
pfcrt	1F	5′-CCGTTAATAATAAATACAGGC-3′	261	NDa	95°C for 5 min; (95°C for 35 s; 56°C for 40 s; 72°C for 1 min) × 35 cycles; 72°C for 10 min
	1R	5′-CTTTAAAAATGGAAGGGTGT-3′
pfmdr1	2F (86–184)	5′-TGTTGAAAGATGGGTAAAGAGCAGAAAGAG-3′	780	ND	95°C for 5 min; (95°C for 40 s; 58°C for 50 s; 72°C for 1 min) × 36 cycles; 72°C for 10 min
	2R (86–184)	5′-TACTTTCTTATTACATATGACACCACAAACA-3′
pfmdr1	3F (1034–1246)	5′-AGAAGATTATTTCTGTAATTTGATACAAAAAGC-3′	880	ND	95°C for 5 min; (95°C for 40 s; 60°C for 42 s; 72°C for 1 min 20 s) × 38 cycles; 72°C for 10 min
	3R (1034–1246)	5′-ATG ATT CGA TAA ATT CAT CTA TAGCAG CAA-3′
Multiplex
pfcrt	K76T F	5′-TGTGCTCATGTGTTTAAACTTAT-3′	134	ApoI	95°C for 5 min; (95°C for 30 s; 56°C for 40 s; 72°C for 1 min) × 32 cycles; 72°C for 10 min
	K76T R	5′-CAAAACTATAGTTACCAATTTTG-3′
pfmdr1	N86Y F	5′-TTTACCGTTTAAATGTTTACCTGC-3′	310	AflIII	95°C for 5 min; (95°C for 35 s; 58°C for 40 s; 72°C for 1 min) × 38 cycles; 72°C for 10 min
	N86Y R	5′-CCATCTTGATAAAAAACACTTCTT-3′
	Y184F F	5′-AAA GAT GGT AAC CTC AGT ATC AAA GAA GAG-3′	560	DraI	95°C for 5 min; (95°C for 35 s; 58°C for 40 s; 72°C for 1 min) × 34 cycles; 72°C for 8 min
	Y184F R	5′-GTC AAA CGT GCA TTT TTT ATT AAT GAC CAT TTA-3′
	S1034C F	5′-AGAATTATTGTAAATGCAGCTTTATGGGGAC TC-3′	230	DdeI	95°C for 5 min; (95°C for 38 s; 60°C for 42 s; 72°C for 1 min 20 s) × 35 cycles; 72°C for 10 min
	S1034C R	5-′AATGGATAATATTTCTCAAATGATAACTTAGCA-3′
	N1042D F	5′-TATGTCAAGCGGAGTTTTTGC-3′	337	AsnI	95°C for 5 min; (95°C for 40 s; 58°C for 40 s; 72°C for 1 min) × 38 cycles; 72°C for 10 min
	N1042D R	5′-TCTGAATCTCCTTTTAAGGAC-3′
	D1246Y F	5′-GTGGAAAATCAACTTTTATGA-3′	499	EcoRV	95°C for 5 min; (95°C for 42 s; 60°C for 40 s; 72°C for 1 min) × 42 cycles; 72°C for 10 min
	D1246Y R	5′-TTAGGTTCTCTTAATAATGCT-3′

^aND, not done.