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We developed a low-cost, ligation-based assay for genotyping single nucleotide polymorphisms (SNPs) in mitochondrial DNA (mtDNA) that will allow inexperienced high school and college students to trace their maternal ancestry. We obtained DNA samples from 155 volunteers and genotyped their mtDNA with the Illumina GoldenGate SNP Genotyping assay. These samples served as standards for the development ... read moreof our simple assay. Our assay was based on the ligation of a biotinylated allele specific probe with a poly-dA tailed locus specific probe such that a positive reaction generated a single DNA strand that was functionalized with both biotin and a poly-dA tail. We developed a simple lateral flow biosensor (LFB) that immobilized this ligation product conjugated with red microspheres resulting in the appearance of a red line on the LFB. The ligation-based assay allowed us to place individuals into haplogroups and thereby determine in what part of the world their maternal ancestors lived. Finally, we performed a case study in which high school students successfully determined their maternal ancestry using our simple assay.read less
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