Pulp was earlier thought to be the major source of DNA in the tooth but the extraction of DNA from the dentin is also on the rise, where the odontoblastic processes are rich in mitochondria which in turn contain mtDNA. Therefore, the study was performed to extract the DNA from odontoblastic process of permanent tooth dentin and to analyze the role of DNA in forensic dentistry. Thereby in this study, permanent maxillary and mandibular premolar teeth, which were extracted for orthodontic reasons, were subject to chemical process to extract the dentin. The extracted dentin was used for PCR analysis to study the DNA and most of the samples inclusive of DNA from both coronal and root dentin samples showed both higher molecular weight band and smear though the concentration of DNA varied among samples. Statistically the mean of total DNA concentration in radicular dentin samples was higher than that of coronal dentin samples. Therefore in our study it is concluded that dentin is a source of DNA. However, radicular dentin is a better source of DNA extraction compared to the coronal dentin.