Human identification analysis to forensic purposes with two mithocondrial markers in polyacrilamide mini gel.

The classical methods for species identification based on protein analysis are not more applicable to the forensic casework. In the last years the analysis of cytochrome b (cyt b) and other mitochondrial markers showed a good feasibility to species determination and individual human identification. The use of cytochrome b is well-known for species detection, even if sequence analysis and comparison in BLAST made this analysis troublesome. In this paper we propose a new method for human sample identification based on polyacrilamide mini gel of a duplex PCR product amplified from mitochondrial DNA corresponding to cytochrome b and a new 16S rRNA fragment that is human-specific and never used to this purpose so far. Multiplex amplification showed a band of 359 bp for the cyt b and a second band of 157 bp for 16S rRNA fragment for human biological samples. Under the same conditions only cyt b fragment was evidenced for sample of animals different from human whereas 16S rRNA fragment was completely absents This method may be very useful for forensic purposes since 16S ribosomal mtDNA fragment is a small human-specific fragment easily amplifiable even with old and highly degraded specimens