The availability of grant funding allowed the OCME-DNA Unit to implement mtDNA processing in order to aid the medical examiner in the identification of remains and support criminal investigations that involve hair evidence (hair shafts or hairs with insufficient roots for nuclear DNA testing). Because of the lack of State/local laboratories with mtDNA capabilities, the OCME-DNA Unit had no “role model” laboratory to follow. The validation plan for mtDNA processing was separated into the following sections: Preliminary Research, Sensitivity and Contamination, ExoSAP-IT Optimization, BigDye Optimization, Sequencing and 310 Optimization, Sample Preparation and Extraction Optimization, Heteroplasmy, Mixtures, and Reproducibility. From the outset of the validation project, one of the few clear goals was to validate a nonorganic extraction method. The advantages would include time efficiency and the elimination of hazardous chemicals from the protocol that is currently used to extract nuclear DNA samples at the OCMR-DNA Unit. The decision was made to use Qiagen QIAamp DNA Blood Mini/Midi columns based upon cost comparison and the ability to remove contaminants and inhibitors efficiently. All sections of the validation were successfully completed, and mtDNA processing of skeletal remains, teeth, and hairs, as well as blood and buccal references samples was adopted by the OCMR-DNA Unit. 5 tables, 3 figures, and 22 references