The first objective of this study was the detection of mitochondrial hypervariable HVII and HVIII regions. Secondly, the study evaluates the importance of these positions for forensic genetic purposes and establishes the degree of variation characteristic of a fragment. Blood samples were collected from 270 healthy unrelated male living in Middle and South of Iraq. FTA® Technology was utilized to extract DNA. A portion of a noncoding region encompassing positions 37 to 340 for HVII and encompassing positions 438 to 574 for HVIII, was amplified in accordance with the Anderson reference sequence. By using EZ-10 spin column the PCR products were purified, sequenced and detected by using the ABI 3730xL Genetic Analyzer. New polymorphic positions 57, 63, 101, 469 and 482 are described that may be very important for forensic identification purpose in the future. This study shows the importance of the adoption of mitochondria in forensic medicine and criminal diagnosis and a private Iraqi society was discovered as the study sites. Further study on larger number of samples from different Iraqi ethics groups is suggested to confirm the results obtained by this study.