Forensic Identification Of 12 Mammals Species Based On Size Variation Of Mitochondrial Cytochrome B Gene Using Multiplex Pcr Assay

In this study, we have applied the endpoint PCR method based on size variation of mitochondrial cytochrome b gene among mammals for identification of 17 mammals. We successfully identify 12 out of 13 mammal species common in the geographic region of Slovenia (Central Europe and West Balkan) using multiplex PCR with our own designed primers for 7 species (cat, pig, lynx, roe and red deer, wolf, bear) and a conventional system, i.e. non-fluorescent primers and agarose gel electrophoresis. Only primer designed for identification of Eurasian lynx turned out to be unspecific. Probably, because it was designed on to sequence data from species close to Eurasian lynx while sequence data for Eurasian lynx was not available in databases at the time of our research. With this method we could not distinguish between subspecies (wolf from dog, and pig from wild boar). The species-specificity of PCR amplicons was further demonstrated by the ability of the assay to accurately identify species-specific DNA from mixed samples. (C) 2011 Elsevier Ireland Ltd. All rights reserved.