ArticleMolecular biology reports2026
Assay for numerical and graphical detection of Native American mitochondrial haplogroup B with real-time PCR.
Article in Molecular biology reports, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
backgroundMitochondrial haplogroup B is characterized by a 9-base-pair deletion (CCCCCTCTA) located between nucleotide positions 8,272 to 8,289 of the human mitochondrial genome. Conventional molecular approaches for detecting this deletion include PCR amplification followed by polyacrylamide gel electrophoresis, or PCR coupled with capillary electrophoresis.
methodsIn this study, we propose a numerical and graphical method based on real-time PCR that combines melting temperature (Tm) determination with melting-curve analysis to distinguish between haplogroup B and non-B individuals.
resultsUsing 1 ng of DNA, samples belonging to haplogroup B exhibited a mean theoretical Tm of 76.1550 °C, whereas non-B sample exhibited a mean Tm of 76.8250 °C. Across different DNA concentrations, the mean experimental Tm of haplogroup B samples differed from that of non-B samples by 0.7258 ± 0.0526 °C. Statistical analysis identified the optimal DNA concentration range for this technique as 0.7-2 ng per real-time PCR reaction. These observations indicate that 0.5-1 ng of DNA per assay yield consistent and reliable results. It is recommended that each assay include minimum one pair of haplogroup B and non-B samples as controls.
conclusionsThe proposed real-time PCR method enables accurate identification of the 9-base-pair deletion characteristics of haplogroup (macrogroup) B, while posing lower health risks, requiring less analysis time and fewer resources than conventional techniques. This approach is applicable in evolutionary, clinical, and forensic investigations.
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