Detection of 484C>G mutation in peroxisome proliferator activated receptor alpha (PPARA) in human DNA using allele-specific polymerase chain reaction

PPARA is encoded by PPARA gene and offers the highest affinity for fatty acids binding. Genetic mutation or polymorphism of this gene in human DNA may results in greater defects on the human normal lipid metabolism. Hence, genetic mutation detection of this gene could be significant in determining the appropriate drug therapy for an individual patient especially for the lipid lowering drug therapy. The purpose of the study is to detect the mutation of 484C>G in PP ARA in human DNA. Previous detection of this gene is done by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) as the detection method. Four samples of human DNA were tested for the detection of the presence or absence of the mutation using an allele-specific polymerase chain reaction (PCR) method in a thermocycler. The results were obtained by running the PCR product under gel electrophoresis using 1.5% of agarose gel. First run of PCR produced amplicons of the DNA of interest while the second run of PCR produced amplicons of both wild type and mutant alleles. All samples were found to be heterozygous as the results showed that the wild type and mutant allele are present.