engleski

Design of species specific primers for amplification of fallow deer (Dama dama) mitochondrial DNA

Species identification based on analysis of short DNA sequences is an essential tool in wildlife management and conservation, in forensic veterinary medicine and meat control in food industry. Control region, a non-coding part of mitochondrial DNA, is a valuable marker for phylogenetic research, population and evolution biology. The goal of our research was to design species specific primers for amplification of fallow deer (Dama dama) control region and to optimize PCR protocol for application of those primers. Primers have been designed based on fallow deer sequences retrieved from GenBank, using Primerblast application for primer design. We have identified 243 base pair PCR product specific for fallow deer. With these species specific primers (F: 5'-TCGCTCCGGGCCCATAGACT-3' ; R: 5'-GCTCCGGGTCGGGGCCTTAG-3') fallow deer can be identified and distinguished from other species using only DNA isolation, PCR (annealing temperature 70° C) and electrophoresis. Application of the primers has been tested on eight samples of fallow deer, three samples of roe deer (Capreolus capreolus) and three reed deer (Cervus elaphus) samples. These species specific primers can be applied in ecological researches, researches of predators feeding habits, in forensic cases and in control of game meat products in food industry.

fallow deer; Dama dama; mitochondrial DNA; control region; primers

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