Fig. 1

Schematic representation of overlap extension PCR to create specific nucleotide at the target site of pvmdr1 gene. To construct a mutant control allele for codon 976 and a wild-type control allele for codon 1076 of pvmdr1 gene, overlap extension PCR was performed. In separate reactions, two overlapping fragments were amplified using Flanking F/Internal R and Internal F/Flanking R primer pairs. The internal primers (Internal F or Internal R) were designed to include the mismatch at 976 or 1076 positions to mimic the mutant or wild-type sequences, respectively. In the second round of PCR reaction, the extension of the overlapped fragments was carried out using Flanking F/Flanking R primer pair