Development of PCR-based method for detection of the polymorphism of n­-methyl-d-aspartate receptor gene / Farrah Idayu Abdul Mutalib

The purpose of this study was to design the allele-specific polymerase chain reaction for detection of the polymorphism ofN-Methyl-D-Aspartate receptor gene and establish the frequencies of the selected genetic variants amongst Chinese ethnic in Malaysia. Single nucleotide polymorphism (SNP) of GRIN2B, rsl805247 was selected to be studied. The allele-specific primers were designed by using Oligo Explorer software and the unique sequence of the primers has been sent to First Base Laboratory Sdn. Bhd to be synthesised. The primers were Rcm_2_NMDAR, Fmt _2_NMDAR and Fwt_2_NMDAR. The most suitable annealing temperature for these primers was found to be 67°C where the intended band appeared and matched with DNA sequencing results. In this study, the most common genotype was GIA (55.8%) and A was the predominant allele (62.8%). It can be concluded that the method has been successfully developed because it matched with gold standard validation method. The assay would be useful for detection of the genetic polymorphism of NMDAR rs1805247 because variations that occurred in analysed samples are more than 1%.