Tetra Primer ARMS PCR Optimization to Detect Single Nucleotide Polymorphism of the KLF14 Gene - Open Access Library Journal

OALibJ > Vol.4 No.12, December 2017

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

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DOI: 10.4236/oalib.1104145 1,769 Downloads 6,883 Views Citations

Author(s)

Ana Sofia Zabala, Miriam Ester Vasquez Gomez, Micaela Fernanda Álvarez, Susana Siewert

Affiliation(s)

Laboratory of Diabetes, Department of Biochemistry and Biological Science, School of Chemistry, Biochemistry and Pharma-cy, National University of San Luis, San Luis, Argentina.

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.

KEYWORDS

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

Share and Cite:

Zabala, A. , Gomez, M. , Álvarez, M. and Siewert, S. (2017) Tetra Primer ARMS PCR Optimization to Detect Single Nucleotide Polymorphism of the KLF14 Gene. Open Access Library Journal, 4, 1-14. doi: 10.4236/oalib.1104145.

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