Development of Tetra-primer ARMS PCR based genotyping method for A clinically important SNP CYP2C19*3 (rs4986893)

Abstract

Treatment of a particular disease with satisfactory outcome has become quite a rare and challenging issue nowadays. This issue is mostly responsible for the polymorphism in the metabolizing gene which alters its metabolized drug function. To solve this problem, SNP genotyping is a great concept. The SNP used in this study rs4986893 present in the gene CYP2C19 causes loss of function to the associated enzyme. As a result, an individual carrying a single copy of the mutant allele becomes a poor metabolizer for a very important anti-coagulant drug, Clopidogrel. The study is conducted by the tetra primer ARMS PCR technique which is an allele specific PCR. The PCR utilize four sets of primer by differentiating homozygous and heterozygous product. Here, in the 3’ end of the primordial the single base mismatch is added. This mutation causes the primordial to amplify a single allele. The ARMS PCR is a cost effective and easy handling procedure. To do the PCR, buccal cells were collected from various range of healthy individual by and their DNA is extracted in phenol-chloroform-isoamylalcohol extraction protocol. The extracted DNA was tested by ARMS PCR method to identify if the individual was carrying the SNP variant or not by resolving the bands in agarose gel electrophoresis. To complete the study with the population perspective of people carrying the SNP variant CYP2C19*3 more thermal cycle optimization step was needed for accomplishing further works.