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Tetra Primer ARMS PCR Optimization to Detect Single Nucleotide Polymorphism of the KLF14 Gene

DOI: 10.4236/oalib.1104145, PP. 1-14

Subject Areas: Diabetes & Endocrinology

Keywords: rs972283 (A/G), KLF14 Gene, Type 2 Diabetes Mellitus

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Abstract

GWAS (genome-wide association studies) have associated Type 2 Diabetes Mellitus with the single nucleotide polymorphism (SNPs) rs972283 (A/G) of KLF14 gene with in a global population. The most common techniques used to analyze SNPs are time consuming, multi-step process and require expensive instruments. Therefore, in order to overcome these problems, we have developed a new, rapid and cost effective T-ARMS PCR assay to genotype rs972283. However, the optimization step can be hardworking and laborious. Hence, we propose to demonstrate and discuss critical steps for its development, in a way to provide useful information. In a first step, we design and validate two specific primer pair for T-ARMS PCR. Later, the amplification conditions were optimized for DNA concentration, annealing temperature, Taq DNA polymerase units and primers concentration. The last one was considered the main interference factor for a correct amplification and appropriate band intensity. Finally, the results obtained by T-ARMS PCR were concordant with sequencing. T-ARMS PCR assay developed in our laboratory for genotyping rs972283 (A/G) of KLF14 gene is time saving and cost-effective compared to the available methods used for SNP studies.

Cite this paper

Zabala, A. S. , Gomez, M. E. V. , Alvarez, M. F. and Siewert, S. (2017). Tetra Primer ARMS PCR Optimization to Detect Single Nucleotide Polymorphism of the KLF14 Gene. Open Access Library Journal, 4, e4145. doi: http://dx.doi.org/10.4236/oalib.1104145.

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