Multiplex polymerase chain reaction (PCR) and fluorescence-based capillary electrophoresis for identification of deer species from antlers

Abstract

Multiplex polymerase chain reaction (PCR) and fluorescence-based capillary electrophoresis (CE) of blood and tissue samples have been used to distinguish between deer species such as red deer, sika deer, wapiti and reindeer. We constructed 4 species-specific primers by using the D-loop of mitochondrial DNA and cytochrome b as an internal PCR control. The amplified species-specific product lengths of 199, 299, 245 and 375 bp for red deer, sika deer, wapiti subspecies and reindeer, respectively were detected from mitochondrial D-loop DNA fragments. The specificity was confirmed by analysis of blood and tissue samples of 4 deer antlers and 8 other samples of mammalian DNA (mouse, rat, pig, chicken, cat, cow, dog and human DNA). The specificity and accuracy of the multiple primers for identification were assessed at various concentrations and from mixed samples. In this study, we established a method for the identification of deer species using deer antlers.

Key words: Deer antler, identification, mitochondrial DNA, multiplex polymerase chain reaction (PCR).