Sessions

The antler of the deer family (Cervidae) is a uniquely fast-growing, annually regenerating bone tissue, which can be an excellent source for genetic studies. The DNA extracted from antlers is useful not only in population genetics and conservation research but can also provide valuable evidence in forensic investigations. However, among the currently described extraction methods from antlers, some require special laboratory equipment or the procedure takes a very long time, others do not contain the necessary information regarding the replicability or the quantity and purity of the extracted DNA, and some may have limitations regarding the type of sample used and the targeted genome region. The aim of our research was therefore to develop a modified protocol that is simpler and faster, while reliably providing a sufficient quantity and quality of DNA for a wide range of genetic studies.

We examined a total of 30 antler samples from roe deer, red deer, and fallow deer individuals (10 samples per species), from which, after a thorough surface cleaning, we obtained a sample by drilling at low rotation speed. After mechanical crushing of the antler samples, four hours of digestion and decalcification followed, then the sample was purified with an organic solvent (phenol–chloroform–isoamyl alcohol), and finally it was concentrated by ultracentrifugation. The quantity and quality of the extracted DNA was verified with agarose gel electrophoresis, Qubit fluorometry, and Nanodrop spectrophotometry. To prove the usability of our method, modified in several aspects compared to the techniques described before, we performed species-specific nuclear microsatellites amplifying multiplex PCR tests.

Based on the parameters examined, no significant differences were observed between the results of the three deer species samples. The examinations aimed at assessing the integrity, concentration, and purity of the extracted genetic material confirmed that the majority of the samples provided sufficient quantity and quality of DNA for downstream applications. At the same time, the samples with suboptimal quantity or purity values also yielded a complete genetic profile by PCR analysis, which supports the applicability of the developed protocol even for nuclear studies.

This procedure, developed based on these results, reliably provides a sufficient amount and quality of DNA even for further genetic analyses, even from small amounts of strongly mineralized samples. Although the protocol should be handled with increased caution as it contains a chemical (phenol) that is hazardous to the health of individuals, it still provides quantities and purity equivalent to the traditional bone-mill and liquid-nitrogen-requiring methods, in a simpler and more cost-effective way. For further testing, we would like to include other calcified sample types (e.g., skull trophies), as well as antler samples of different ages from other species of deer.