Abstract
Molecular sexing of wild animals is feasible with differential amplification of X- and Y-homologous regions of various genes. We present a simple and robust technique for sex identification of red panda suitable for noninvasive studies based on sex-specific fragment size polymorphism in amelogenin (AMEL) gene. Sequence analysis of AMELX and AMELY revealed an 18-bp deletion in Y fragment which was used to differentially amplify 217 bp of X and 199 bp of Y fragments. The designed primer pair is very sensitive and exhibits polymerase chain reaction (PCR) success rates of 89 and 78 % at 0.1 and 0.01 ng template DNA concentrations, respectively. Further, we amplified poor quality faecal DNA with 94 % success rate and average dropout rate of 15.6 %. In silico analysis shows that these primers can also be used to identify sex of a sympatric species, giant panda.
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Acknowledgments
We gratefully acknowledge Mr. Alankar Jha, Director of Padmaja Naidu Himalayan Zoological Park, Darjeeling, India, and Dr S. Shivaji for taking a keen interest and supporting this work.
Funding
This study was funded by the Central Zoo Authority, Government of India.
Conflict of interest
The authors declare that they have no competing interest.
Ethical approval
Blood samples used in this study were acquired from the Padmaja Naidu Himalayan Zoological Park, Darjeeling, India, following the Padmaja Naidu Himalayan Zoological Park and Central Zoo Authority, Government of India, ethical guidelines and procedures. No animals were killed or captured as a result of this study.
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Communicated by M. Scandura
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Fig. S1
Alignment showing Amelogenin sequence polymorphism in X- and Y-homologous fragments in giant panda and red panda (GIF 201 kb)
Fig. S2
Agarose gel (2.5 %) electrophoresis of Amelogenin locus in female (a), and male (b) red pandas with serial dilutions of blood DNA. M – 100 bp size marker, N – negative control (GIF 342 kb)
Fig. S3
Agarose gel (2.5 %) electrophoresis of Amelogenin locus with faecal DNA of captive red pandas. M – 100 bp size marker, N – negative control, ♀ - female, ♂ - male (TIFF 2288 kb)
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Kumar, A., Roka, B., Rai, U. et al. Molecular sex identification of red panda (Ailurus fulgens) suitable for noninvasive genetic studies. Eur J Wildl Res 61, 641–644 (2015). https://doi.org/10.1007/s10344-015-0928-2
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DOI: https://doi.org/10.1007/s10344-015-0928-2