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Powder-Free DNA Extraction from Post-Mortem Teeth

Abstract

Heitor Simoes Dutra Correa, Lucas Gabriel Sena Carneiro, Nelson Massayuki Yoshitake, Andrea Correia Carneiro and Geter Sinear Jesus Bizo

In order to perform human identification through DNA analysis, bones and teeth are considered the samples with the highest success rates. Regarding the protocols used to process these samples, they are also usually submitted to the same treatment, which consists of sample powdering. However, there are tissues in a tooth that have low DNA content, and thus, should not be included in DNA extraction. Recent studies have reported that sampling of dental cementum could optimize DNA typing from teeth due to its high cellularity and resilience. The objective of this article is to present an improved protocol for DNA extraction from teeth that targets cementum and its use in two real cases. After a 24-hour decalcification step, cementum was sampled and submitted to a simple DNA extraction protocol based on a commercial kit and automated platform. Real-time PCR (Polymerase Chain Reaction) quantification, PCR amplification of 27 DNA markers and capillary electrophoresis were performed. For both real cases, a different sample type had yielded a full DNA profile using a different and validated protocol. Quantification showed expected concentration and integrity of DNA in both tooth samples. After genotyping, all samples presented full and concordant profiles. This pilot study demonstrated that the presented protocol, specifically designed for teeth, was able to obtain full DNA profiles from a burnt and a skeletonized human body.

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