Resumo

SIERRA, Wilson Uriel; DEL-CASTILLO-SABOGAL, Hernando  e  ESPITIA-ORTIZ, Tatiana. Unified digestion and decalcification protocol for three DNA extraction methods in human remains. Rev. acad. colomb. cienc. exact. fis. nat. [online]. 2017, vol.41, n.161, pp.447-455. ISSN 0370-3908.  https://doi.org/10.18257/raccefyn.527.

The identification of human remains using DNA from bones and teeth is a current procedure in forensic laboratories worldwide. At least three methods are used in DNA extraction from mineralized tissue: Organic (phenol-chloroform), silica (QIAquick, QIAgen) and magnetic particles (Prepfiler Express™ BTA Forensic DNA Extraction Kit); each one has its own procedure for decalcification, digestion, DNA purification, and elution, as well as for obtaining the initial amount of material. Although the three methods are equally efficient, we standardized a decalcification- digestion protocol to use it with any of the three DNA purification methods. We evaluated the quantity of powder, detergent and protease concentration, device concentrators and buffer for demineralization in bones and teeth from forensic cases of the Laboratory and DNA of common cellular lines by assessing the amount of recovered DNA and the quality of the genetic profile. The unified protocol for digestion and decalcification allowed a full degradation of bone powder, and good quality DNA recovery, independently of the purification method used in the subsequent step. Success percentage with this protocol was nearly 80 %.

Palavras-chave : Human remains; Digestion-decalcification; DNA extraction; PrepFiler Express™; Silica; Phenol-chloroform.

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