GENETIC CHARACTERIZATION OF CERTAIN CAPTIVE WILD CARNIVORES USING RANDOM-AMPLIFIED POLYMORPHIC DNA-POLYMERASE CHAIN REACTION
2025
B. Nishanth | C. Sreekumar | K.G. Tirumurugaan | G. Sarathchandra | K. Senthil Kumar
Wildlife species identification, based on DNA is a highly reliable method for the investigation of wildlife crimes. PCR techniques and microsatellite markers are extensively used for the identification of wild animal DNA in forensic cases. Mostly, genes like Cyt B, Cox 1 and 12S RNA are amplified coupled with sequencing, to confirm the species in question. In the present study, random amplified polymorphic DNA (RAPD) PCR was performed with DNA samples from captive wild felids, canids and ursids and the finger-print pattern was analyzed for their utility in designing de novo diagnostic primers. DNA was extracted from the tissue samples from eleven animals (two numbers each from tigers and lions and one sample each from a leopard, jaguar, sloth bear, black bear, wolf, jackal and dhole). RAPD-PCR was carried out using different arbitrary decamers. Of the five decamers used, two decamers, namely AP7 and AP17 revealed consistent amplification patterns. Among them, AP7 produced monomorphic fragments corresponding to 500 bp for felids and 800 bp and 400 bp for ursids, while polymorphic bands were generated across the different genera. The decamer AP17 consistently amplified a 450 bp band from all the felid samples and polymorphic amplification with other genera. The potential for these RAPD amplicons for their applicability in the design-specific primers for genus/ species-specific detection of wild animal DNA due to their uniqueness is discussed.
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