The evidential value of an mtDNA match between biological remains and their potential donor is determined by the random match probability of the haplotype. This probability is based on the haplotype’s population frequency estimate. Consequently, implementing a population study representative of the population relevant to a forensic case is vital to correctly evaluating the evidence. The emerging number of poaching cases and the limited availability of such data emphasizes the need for an improved fallow deer mtDNA population databank for forensic purposes, including targeting the entire mitochondrial control region. By sequencing a 945-base-pair-long segment of the mitochondrial control region in 138 animals from five populations in Hungary, we found four different haplotypes, including one which had not yet been described. Our results, supplemented with data already available from previous research, do not support the possibility of determining the population of origin, although some patterns of geographical separation can be distinguished. Estimates of molecular diversity indicate similarly low mtDNA diversity (Hd = 0.565 and π = 0.002) compared to data from other countries. The calculated random match probability of 0.547 shows a high probability of coincidence and, therefore, a limited capacity for exclusion. Our results indicate that despite the overall low genetic diversity of mtDNA within the Hungarian fallow deer samples, a pattern of differentiation among the regions is present, which can have relevance from a forensic point of view.

The fallow deer (Dama dama) represents significant game management value globally, and human activities are significantly impacting the species. Besides the positive effects, these activities can threaten its existence, health, and value. The aim of the authors was to develop a tetranucleotide microsatellite panel that could be clearly interpreted and used for genetic testing of fallow deer. Such a panel did not exist until now and could be particularly useful in the field of conservation genetics and forensics. A total of 99 tetrameric microsatellites, originally designed for related deer species, were tested on 20 fallow deer individuals from five Hungarian sampling areas. Original and newly designed primers were used to amplify the microsatellite regions using previously published or optimized PCR protocols. The lengths and sequences of specific amplicons were detected using capillary electrophoresis, and the rate of polymorphism was determined. Altogether, 80 markers provided PCR products of adequate quality and quantity. Among them, 15 markers proved to be polymorphic (2–5 alleles/locus), and 14 tetrameric markers were selected for further analysis. Statistical calculations showed that the selected polymorphic microsatellites can potentially enable key individualization in many areas of wildlife and population genetics, thus protecting the species.

A simple, fast, and cost-effective organic solvent-based protocol was developed for DNA extraction from deer antlers and prepared trophy skulls, eliminating the need for commercial kits or cryogenic grinding. The method combines bead-based mechanical homogenization with a 4 h enzymatic digestion in EDTA buffer containing N-lauryl sarcosine and Proteinase K, followed by phenol–chloroform–isoamyl alcohol purification and centrifugal filtration. DNA quality and quantity were evaluated using agarose gel electrophoresis, Qubit fluorometry, and Nanodrop spectrophotometry. The protocol was tested on 60 samples, comprising 30 antlers and 30 pedicle parts from prepared trophy skulls of roe deer (Capreolus capreolus), fallow deer (Dama dama), and red deer (Cervus elaphus). To assess suitability for downstream applications, species-specific microsatellite markers were amplified using multiplex PCR, successfully generating complete genotypes from all 60 samples. These results, along with a demonstrated case study, confirm that the developed protocol provides high-quality DNA suitable for molecular genetic investigations, enabling reliable genotyping from small amounts of both antler and processed trophy materials in forensic and conservation contexts.