Some biological samples cannot be genotyped with routine STR analyzes, if they contain trace amount of DNA. At these cases, mitochondrial DNA analyzes can be carried out. The aim of this work was optimizing the sequencing of mtDNA's HV1 and HV2 regions from samples like hair, nails, earrings, toothbrushes, q-tips, glass edge swabs, gums, razors and cigarette butts. At this work 5 unrelated individual's samples mentioned above were sequenced and other 11 unrelated individual's blood samples were sequenced for the reliability of the procedure. In this examined Turkish population, most common polymorphic positions were 263, A. >. G point mutation, and 315.1 C insertion. The HV2 profile "263, A-G 315.1 C" observed on 3 individuals and "152, T-C 263, A-G 309.1 C 309.2 C 315.1 C" profile observed on 2 individuals. Two individual also shared a profile same as rCRS on their HV1 regions. Every analyzed person has unique HV1. +. HV2 profiles together. © 2011 Elsevier Ireland Ltd.