Two 16S rRNA mitochondrial markers for species identification in forensic science

In the recent years, with the development of molecular biology technology, new methods for human identification to forensic purposes, based on genetic differences among the animal species have been proposed. The analysis of cytochrome b (cyt b) showed a good feasibility to species determination and individual human identification. The use of cytochrome b is well-known for species detection, even if sequences analysis and comparison in BLAST made this analysis troublesome in case of degraded samples from which it is difficult obtain a good sequence. In this paper we propose a method for the identification of human samples based on the amplification of a duplex PCR corresponding to two 16S mitochondrial fragments, an universal fragment of 236 bp and a human-specific fragment of 157 bp. A rapid analysis of the PCR products can be performed in a mini poliacrylamide gel. The primers for both 16S rRNA fragments were designed by us. Since these two 16S rRNA fragments are small, they amplify easily even if the presence of old and highly degraded specimens in a single round PCR: for these reasons, this method could result very useful for forensic purposes in case of human species identification. (C) 2011 Elsevier Ireland Ltd. All rights reserved.