Automatable full demineralization DNA extraction procedure from degraded skeletal remains

Amory, Sylvain; Huel, René; Bilić, Ana; Loreille, Odile; Parsons, Thomas J.

doi:10.1016/j.fsigen.2011.08.004

Cited by 105 publications

(57 citation statements)

References 15 publications

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“…The samples were then washed (NaClO, purified H 2 O, absolute EtOH), dried, and ground using a horizontally oscillating ballmill. Bone powder was dissolved in a hybridisation oven, the DNA extracted with a modified silica-column-based Spin Filter (SF) method [27, 28]. …”

Section: Methodsmentioning

confidence: 99%

Three individuals, three stories, three burials from medieval Trondheim, Norway

et al. 2017

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This article presents the life stories of three individuals who lived in Trondheim, Norway, during the 13th century. Based on skeletal examinations, facial reconstructions, genetic analyses, and stable oxygen isotope analyses, the birthplace, mobility, ancestry, pathology, and physical appearance of these people are presented. The stories are discussed within the relevant historical context. These three people would have been ordinary citizens, without any privileges out of the ordinary, which makes them quite rare in the academic literature. Through the study of individuals one gets a unique look into the Norwegian medieval society.

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Section: Methodsmentioning

confidence: 99%

Three individuals, three stories, three burials from medieval Trondheim, Norway

et al. 2017

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“…For the DNA extraction from the demineralization/lysis supernatants three published protocols either directly using the silica columns and buffer set included in the QIAamp DNA Blood Maxi kit (Qiagen, Hilden, Germany; SC method) [7], phenol/chloroform/isoamyl-alcohol (PCI) [8], or a spin filter method (SF) [9] were compared in a series of initial experiments on three specimens. Both the PCI and the SF protocol featured a final purification step on a silica matrix using the MinElute PCR Purification kit (Qiagen) according to the manufacturer's recommendations with two additional washing steps.…”

Section: Methodsmentioning

confidence: 99%

“…This additional step in the protocol appeared to attenuate the risk of filter-clogging and reduced the concentrations of the lysis buffer constituents in the retenate recovered from the spin filters. All further steps of the SF protocol were performed as described in [9]. The final volume of DNA extracts was 50–75 μl for all approaches.…”

Section: Methodsmentioning

confidence: 99%

Comparison of morphological and molecular genetic sex-typing on mediaeval human skeletal remains

Bauer

Niederstätter

McGlynn

et al. 2013

Forensic Science International: Genetics

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Archaeological excavations conducted at an early mediaeval cemetery in Volders (Tyrol, Austria) produced 141 complete skeletal remains dated between the 5th/6th and 12th/13th centuries. These skeletons represent one of the largest historical series of human remains ever discovered in the East Alpine region. Little historical information is available for this region and time period. The good state of preservation of these bioarchaeological finds offered the opportunity of performing molecular genetic investigations. Adequate DNA extraction methods were tested in the attempt to obtain as high DNA yields as possible for further analyses. Molecular genetic sex-typing using a dedicated PCR multiplex (“Genderplex”) gave interpretable results in 88 remains, 78 of which had previously been sexed based on morphological features. We observed a discrepancy in sex determination between the two methods in 21 cases. An unbiased follow-up morphological examination of these finds showed congruence with the DNA results in all but five samples.

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“…Con base en los experimentos previos en el Laboratorio, así como en reportes de la literatura (Loreille, et al, 2007;Amory, et al, 2012), se partió de 1 g de material pulverizado para evaluar la eficiencia de la digestión, se utilizó un volumen de 15 ml de solución tampón para digestión compuesto de EDTA (JT Baker), lauril sarcosinato de sodio (LSS) (Sigma-Aldrich) y 10mg/ml de proteinasa K (pK) (AG Scientific).…”

Section: Materiales Y Métodosunclassified

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

Sierra¹,

del-Castillo-Sabogal²,

Espitia-Ortiz³

2018

Rev. Acad. Colomb. Cienc. Ex. Fis. Nat.

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ResumenLa identificación de restos humanos esqueletizados con base en el ADN es un procedimiento muy utilizado en los laboratorios forenses del mundo. Existen al menos tres métodos para la obtención del ADN a partir de este tipo de tejido: mediante solventes orgánicos (fenol-cloroformo), columnas de sílice (QIAquick de QIAgen) y perlas magnéticas recubiertas de polímero (PrepFiler Express™ BTA Forensic DNA Extraction Kit). Cada uno de ellos involucra un procedimiento distinto para la digestión y descalcificación del tejido óseo, así como la aplicación de diferentes principios fisicoquímicos para la purificación y la elución del ADN y el uso de diferentes cantidades iniciales de material pulverizado. Aunque los tres métodos han demostrado ser efectivos dependiendo de la calidad de la muestra, en el Laboratorio de Investigación Genética de Restos Humanos del Instituto Nacional de Medicina Legal y Ciencias Forenses se estandarizó un protocolo unificado para la digestión y descalcificación, independientemente del método de purificación. Con este fin, se evaluaron variables como la cantidad de material pulverizado, el uso y la concentración de detergentes, la concentración de proteinasa K, el uso de dispositivos concentradores y de los reactivos del estuche PrepFiler Express™ BTA Forensic DNA Extraction y EDTA. El efecto de las variables se evaluó en muestras de tejido calcificado provenientes de los casos forenses del Laboratorio y en el ADN de líneas celulares conocidas analizando la cantidad de ADN recuperado y la calidad del perfil genético. El protocolo unificado de digestión y decalcificación permite una digestión completa del material pulverizado y, por ende, una mejor recuperación de ADN, independientemente del método de purificación usado. En las muestras analizadas el método tuvo un porcentaje de éxito cercano a 80 %. © 2017. Acad. Colomb. Cienc. Ex. Fis. Nat. Palabras clave:Restos óseos; Digestión-decalcificación; Extracción de ADN; PrepFiler Express™; Sílice; Fenol-cloroformo. Abstract Unified digestion and decalcification protocol for three DNA extraction methods in human remainsThe 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 decalcificationdigestion 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. ...

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Automatable full demineralization DNA extraction procedure from degraded skeletal remains

Cited by 105 publications

References 15 publications

Three individuals, three stories, three burials from medieval Trondheim, Norway

Three individuals, three stories, three burials from medieval Trondheim, Norway

Comparison of morphological and molecular genetic sex-typing on mediaeval human skeletal remains

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

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