Study on insulin gene rs689 polymorphism by means of allele-specific polymerase chain reaction

Abstract. \n\nAim. The work was initiated to develop a modified protocol for PCR method to determine INS gene rs689 polymorphism.\n\nMaterials and methods. To study the INS gene rs689 polymorphism, we used a modified method of allele-specific PCR. Original design of primers was performed by means of bioinformation analysis of NCBL database, Genome Browser with BioEdit app. The PCR product was analyzed by electrophoresis in 3 % agar gel.\n\nResults and Discussion. We have managed to develop and optimize the allele-specific PCR analysis to study the insulin gene rs689 polymorphism. As the method requires no high qualification of the staff and high-tech expensive equipment but is suitable for automation, it is advantageous of similar others. Analysis of results from genotyping of INS gene rs689 polymorphism in patients with diabetes mellitus helped identify association of T allele and heterozygous A/T genotype with the risk of DM onset.\n\n Conclusion. The modified protocol based on the allele-specific PCR intended for study on the INS gene rs689 polymorphism was developed. The allele-specific PCR allows direct diagnosis of some genetically determined disorders without additional steps, to name hybridization, ligation or endonuclease restriction. The procedure is simple and accurate, and could be a cost-effective alternative to highly efficient specific primers significantly simplifying the studies.

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