TITLE:
Designing, Optimization and Validation of Tetra Primer ARMS-PCR Protocol for Genotyping Single Nucleotide Polymorphism rs4731702 (C/T) of KLF14 Gene Associated with Type 2 Diabetes Mellitus: A Study in San Luis, Argentina
AUTHORS:
Micaela Fernanda Alvarez, Miriam Ester Vasquez Gomez, Susana Siewert
KEYWORDS:
Molecular Markers, SNP, Insulinemia, Metabolic Syndrome, Dyslipidemias
JOURNAL NAME:
Open Access Library Journal,
Vol.3 No.12,
December
16,
2016
ABSTRACT: New, quick, and inexpensive methods for genotyping KLF14 gene polymorphism through Tetra Primer ARMS-PCR were developed in the present investigation. Single nucleotide polymorphism (SNP) genotyping needs to be attempted to establish association between the identified SNP with metabolic trait for identification of molecular markers that can be used to identify individuals at risk of developing Diabetes Mellitus Type 2. In the current study, we have successfully genotyped the SNP rs4731702 (C/T) in KLF14 gene. Tetra primer ARMS-PCR protocol was optimized and validated for this SNP with short turn-around time and costs. The optimized techniques were tested on 60 samples of controls and type 2 Diabetes Mellitus individuals. Samples with known genotypes for the described gene, previously tested in duplicate using the sequencing methods, were employed for validation of the assay. Upon validation, complete concordance was observed between the Tetra Primer ARMS-PCR assays and the sequencing results. These results highlight the ability of Tetra Primer ARMS-PCR in genotyping of SNP in KLF14 gene. Our results provide direct evidence that Tetra Primer ARMS-PCR is a rapid, reliable, and cost-effective method for SNP genotyping of KLF14 gene in type 2 Diabetes Mellitus individuals.