The objective of this study was species identification of the following biological trace material: skin, blood stains, meat samples and jawbone with a tooth, which were the subject of expert opinion ordered by a court. The expert appraisement was conducted by an analysis of a cytochrome b fragment. The choice of mtDNA fragment for analysis was based on its conservation in mammals which enabled several farm and wild species to be identified with one pair of primers. The PCR product was differentiated by Tsp509I and Alulenzymes. Due to problems with amplification of roe deer DNA, primers specific to this species only, flanking a cytochrome b fragment (Y1495 1.1), were designed. On the basis of this analysis, it was concluded that the skin sample was derived from a goat, dried blood from a roe deer, the jawbone from cattle, and two meat samples from a roe deer and red deer. This method allowed rapid and efficient identification of several species of mammals using diverse biological material.