Figure 2

PCR-RFLP of the pfdhfr gene, amplified region of M4-F primers. The 326 bp fragment was cut by Alu I into 180 and 118 bp fragments, indications for wild type ser108 and 299 bp for mutant (A). BstNI digested only mutant allele into 145 and 181 bp indicates mutation 108thr. All the tested samples showed wild type (B). Reaction with XmnI produced fragment 163 and 189 bp for wild type; and 26, 137 and 163 bps for 59arg mutation (C). Lane L: DNA ladder of Agilent DNA 1000 Kit (Agilent Technologies, Molecular Probes Inc, USA), The controls are in Lane 1 and 2: A: Lane 1: Plasmodium falciparum KI strain (mutant); Lane 2: P. falciparum T9.96 strain (wild type); Lane 3–9: samples from Kalabakan; Lane 10: PCR negative control (no DNA was added to the PCR reaction). B: Lane 1: Plasmodium falciparum KI strain (wild type); Lane 2: P. falciparum T9.96 strain (wild type); Lane 3–9: samples from Kalabakan; Lane 10: PCR negative control (no DNA was added to the PCR reaction). C: Lane 1: Plasmodium falciparum KI strain (mutant); Lane 2: P. falciparum T9.96 strain (wild type); Lane 3–9: samples from Kalabakan; Lane 10: PCR negative control (no DNA was added to the PCR reaction).