Since 1996, the number of individuals performing mitochondrial DNA analysis at the FBI Laboratory has grown from 4 to 12. Over 15 mitochondrial DNA cases have been completed by the Lab as of March 1999. Mitochondrial DNA sequencing is becoming known not only as an exclusionary tool but also as a complementary technique for use with other human identification procedures. Mitochondrial DNA differs from nuclear DNA in its location, its sequence, its quantity in the cell, and its mode of inheritance. The current forensic analysis of mtDNA (contained in mitochondria) is rigorous and labor-intensive. The steps of the mtDNA analysis process include primary visual analysis, sample preparation, DNA extraction, polymerase chain reaction (PCR) amplification, postamplification quantification of the DNA, automated DNA sequencing, and data analysis. Mitochondrial DNA sequences are initially generated by computer software and subsequently edited by a DNA examiner using multiple runs from the same template to obtain the final sequence of the DNA. Mitochondrial DNA analysis will continue to be a powerful tool for law enforcement officials in the years to come This paper presents an overview of mitochondrial DNA analysis at the FBI Laboratory including the background of mitochondrial DNA, analysis procedures, interpretation guidelines, population database, statistics reporting, quality assurance, and testimony. References and figures