Table 2.

PCR programs for PfATP6 codons 263, 623 and 769

PfATP6 Codon	PCR	Temp.	Time	N. of cycles	Primers
263	1st round PCR	92 °C	3 min	1 cycle	263P1 and 263P2
		92 °C	30 sec	30 cycles
		47 °C	45 sec
		65 °C	1 min
		68 °C	5 min	1 cycle
		4 °C	Hold	1 cycle
	2nd round PCR	95 °C	3 min	1 cycle	263WT and 263 N1* 263 M and 263 N1
		95 °C	45 sec	20 cycles
		62 °C	1 min
		72 °C	1 min
		72 °C	10 min	1 cycle
		4 °C	Hold	1 cycle
623-769	1st round PCR	94 °C	5 min	1 cycle	ATP1 and ATP2
		94 °C	30 sec	45 cycles
		44 °C	45 sec
		68 °C	1 min
		68 °C	5 min	1 cycle
		4 °C	Hold	1 cycle
623	2nd round PCR	94 °C	5 min	1 cycle	623WT and 623 N2* 623 M and 623 N2
		94 °C	30 sec	20 cycles
		40 °C	45 sec
		68 °C	1 min
		70 °C	5 min	1 cycle
		4 °C	Hold	1 cycle
769	2nd round PCR	94 °C	5 min	1 cycle	769 N1 and 769 N2
		94 °C	30 sec	25 cycles
		56 °C	45 sec
		68 °C	1 min
		68 °C	5 min	1 cycle
		4 °C	Hold	1 cycle

(*) Second round PCR for codon 263 and 623 were performed in single reactions with either the wild type (WT) or mutant (M) primer.