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Rapid, Inexpensive Genotyping and Barcoding of Primates: Multiple Applications for High-Resolution Melt Analysis in Primatology Open Access

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Research in molecular ecology and conservation genetics often entails genotyping single nucleotide variants (SNVs). High-Resolution Melt Analysis (HRMA) is a simple and economical method for detecting DNA variants by characterizing the sequence-specific melting behavior of short PCR products. To-date HRMA use has largely focused on medical screenings, but this method has numerous potential applications in primatological genetics. We developed and tested (via Sanger sequencing) several protocols demonstrating the convenience and flexibility of HRMA in primatology. First, we assessed the ability of HRMA to discern color vision status in lemurs (n=87 of 9 species) via X-linked opsin genotypes. Differences in melting curves (temperature and shape) allowed us to reliably identify trichromatic and dichromatic individuals with high accuracy. Second, we targeted SNVs commonly associated with autism and/or behavioral tendencies (oxytocin receptor gene, OXTR) in humans (n=60) and were able to accurately genotype individuals based on melting curves. Third, we used HRMA for rapid species identification using a segment of cytochrome c oxidase 1 (COX1). Results indicate that sympatric primate species, including some lemurs and apes, can be accurately identified using HRMA. Finally, we have used HRMA for health screenings of interleukin-4 (IL4) SNVs that are associated with nematode infection loads in Eulemur rufifrons. Our results demonstrate that HRMA is a multipurpose and robust method for genotyping simple functional and neutral genetic variants.

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