Optimization of PCR method for detection of single nucleotide polymorphism of IFIH1 gene / Suhaila Abdul Rahim

IFIH1 is one of the Non-HLA gene that involved in the coding of the immune system. Though not directly involved as the cause of the disease, it was involved in the destruction of the beta-cell of the pancreas due to the altered immune cell system. The method used to detect the allele of interest which was rs3741537 in the IF/HJ gene was executed through the Nested PCR method. This study altogether had mainly focusing on the optimization of the Nested PCR method itself in which different parameters which includes concentration of DNA template, annealing temperature, number of cycle and the volume of primers were altered and experimented with. Upon completion of the Nested PCR, the band formation of the gene was evaluated by the gel electrophoresis. In conclusion, the study in developing the optimization of Nested PCR was successful in which the wild type allele and the mutant type allele was able to be differentiated. Thus, from this study, it can be seen that Nested PCR was a method that can be applied for Malaysian in detecting the possible chances of having the type 1 Diabetes and at the same time also being able to be the possible target for type 1 Diabetes treatment.