Species Determination using Species-discriminating PCR-RFLP of Ancient DNA from Prehistoric Skeletal Remains

Abstract

Interspecific sequence polymorphisms in the mitochondrial cytochrome b gene were analyzed by PCR-RFLP to determine the species origin of Bronze Age animal skeletal remains. Existing techniques were refined by targeted primer design focusing on a DNA fragment shorter than 200 bp, an approach allowing us to identify up to six animal species at the same time. Possible contaminants, such as human DNA, were reliably ruled out. For routine applications in archaeometry, food or material analyses, PCR-RFLP may thus provide a simple alternative to sequencing of PCR products, allowing discrimination between species, even if the template DNA is degraded or contains traces of DNA from various species.