Fig. 1

PCR-RFLP results confirmation using Sanger Sequincing: The figure illustrates the validation of SIRT6 SNP rs117385980 (C > T) genotyping results obtained by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) against Sanger sequencing. Panels (a), (b), and (c): Sanger sequencing chromatograms confirming selected genotypic groups: Panel (a): Shows the heterozygous CT genotype (sample Code: 1707) with overlapping blue (C) and red (T) peaks at the SNP position. Panels (b) and (c): Show the homozygous wild-type CC genotype (samples Code: 1414 and 1694), where only the blue peak (C) is present at the SNP position. Panel (d): Representative PCR-RFLP results separated on a 4% agarose gel. The target fragment was 661 bp prior to restriction digestion. Digestion was performed with the BsuRI (HaeIII) restriction enzyme: CC Genotype (Wild-Type): The amplicon is digested, yielding observable fragments of 241 bp, 216 bp, and 161 bp. (the 43 bp fragment is not discernible on this gel). CT Genotype (Heterozygous): Both the digested wild-type fragments (e.g., 161 bp) and the 402 bp fragment (indicative of the undigested mutant T allele) are visualized. 1 kb: Molecular weight marker (Ladder).