Protocolo unificado de digestión y descalcificación de tres métodos de extracción de ADN de restos humanos

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 %. © 2017. Acad. Colomb. Cienc. Ex. Fis. Nat.

[1]  M. Wasim,et al.  Evaluation of different methods for DNA extraction from human burnt bones and the generation of genetic profiles for identification , 2017, Medicine, science, and the law.

[2]  P. Wiegand,et al.  As solid as a rock—comparison of CE- and MPS-based analyses of the petrosal bone as a source of DNA for forensic identification of challenging cranial bones , 2017, International Journal of Legal Medicine.

[3]  M. Nagy,et al.  Comparison of two silica-based extraction methods for DNA isolation from bones. , 2016, Legal medicine.

[4]  L. Orlando,et al.  Comparing the performance of three ancient DNA extraction methods for high‐throughput sequencing , 2016, Molecular ecology resources.

[5]  D. Higgins,et al.  Teeth as a source of DNA for forensic identification of human remains: a review. , 2013, Science & justice : journal of the Forensic Science Society.

[6]  Pedro Alberto Barrio-Caballero Revisión de métodos de extracción de ADN a partir de restos óseos en el laboratorio forense , 2013 .

[7]  P. Wojtkiewicz,et al.  Technical note: Bone DNA extraction and purification using silica-coated paramagnetic beads. , 2012, American journal of physical anthropology.

[8]  Jason Y. Liu,et al.  AutoMate Express™ Forensic DNA Extraction System for the Extraction of Genomic DNA from Biological Samples * , 2012, Journal of forensic sciences.

[9]  Sylvain Amory,et al.  Automatable full demineralization DNA extraction procedure from degraded skeletal remains. , 2012, Forensic science international. Genetics.

[10]  Eske Willerslev,et al.  DNA in ancient bone - where is it located and how should we extract it? , 2012, Annals of anatomy = Anatomischer Anzeiger : official organ of the Anatomische Gesellschaft.

[11]  J. Jakubowska,et al.  Comparison of three methods of DNA extraction from human bones with different degrees of degradation , 2011, International Journal of Legal Medicine.

[12]  L. Bonewald,et al.  The Amazing Osteocyte , 2010, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[13]  Takahide Yokoi,et al.  Evaluation of a new experimental kit for the extraction of DNA from bones and teeth using a non-powder method. , 2010, Legal medicine.

[14]  S. Zeni,et al.  Marcadores bioquímicos del remodelamiento óseo. Utilidad clínica , 2009 .

[15]  M. Furtado,et al.  Developmental Validation of the PrepFiler™ Forensic DNA Extraction Kit for Extraction of Genomic DNA from Biological Samples * , 2009, Journal of forensic sciences.

[16]  T. Parsons,et al.  Highly effective DNA extraction method for nuclear short tandem repeat testing of skeletal remains from mass graves. , 2007, Croatian Medical Journal.

[17]  T. Parsons,et al.  Success rates of nuclear short tandem repeat typing from different skeletal elements. , 2007, Croatian medical journal.

[18]  T. Parsons,et al.  High efficiency DNA extraction from bone by total demineralization. , 2007, Forensic science international. Genetics.

[19]  N. Rohland,et al.  Comparison and optimization of ancient DNA extraction. , 2007, BioTechniques.

[20]  Suni M. Edson,et al.  Naming the Dead - Confronting the Realities of Rapid Identification of Degraded Skeletal Remains. , 2004, Forensic science review.

[21]  Andrew R. Millard,et al.  The survival of organic matter in bone: a review , 2002 .