A DNA extraction method of small quantities of bone for high-quality genotyping

Autores
Caputo, Mariela; Irisarri, Maximiliano; Alechine, Evguenia; Corach, Daniel
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
DNA genotyping techniques have been used successfully in forensic science for almost three decades and represent the gold standard for individual identification. However, efficient protocols for obtaining DNA from exhumed bones suitable for genotyping are still scarce and most of them require a considerable amount of starting material, are time consuming and are inefficient for reducing inhibitor's effects. We sought to develop an optimised protocol for extracting DNA from bone samples obtained from exhumations. We tested two approaches for preparing bone samples: (a) fine powder and (b) thin slices of bone. The best ratio of bone amount to DNA yields was assessed by a titration experiment using bone powder ranging from 50 to 1000mg. We obtained optimal DNA yields (27pgmg(-1) on average) when 150-200mg of starting material were processed using a one-step demineralisation method. Better-quality profiles (determined by the number of genotyped loci) were obtained when DNA was extracted from bone slices compared to extraction from bone powder. From bone slices 83.9% and from bone powder 46.7% of the samples provided genotypes for 11 or more loci. Since bone preparation procedures were carried out at room temperature, the method developed in the present study might be an attractive alternative to the standard freeze-mill approach, being faster and more cost-efficient.
Fil: Caputo, Mariela. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina;
Fil: Irisarri, Maximiliano. Universidad de Buenos Aires. Facultad de Farmacia y Bioquimica. Servicio de Huellas Digitales Geneticas;
Fil: Alechine, Evguenia. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina;
Fil: Corach, Daniel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina;
Materia
Exhumed-Bone
Dna Extraction
Expert Software
Str Profile
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/1992
Acceder
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network_name_str CONICET Digital (CONICET)
spelling A DNA extraction method of small quantities of bone for high-quality genotypingCaputo, MarielaIrisarri, MaximilianoAlechine, EvgueniaCorach, DanielExhumed-BoneDna ExtractionExpert SoftwareStr Profilehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1DNA genotyping techniques have been used successfully in forensic science for almost three decades and represent the gold standard for individual identification. However, efficient protocols for obtaining DNA from exhumed bones suitable for genotyping are still scarce and most of them require a considerable amount of starting material, are time consuming and are inefficient for reducing inhibitor's effects. We sought to develop an optimised protocol for extracting DNA from bone samples obtained from exhumations. We tested two approaches for preparing bone samples: (a) fine powder and (b) thin slices of bone. The best ratio of bone amount to DNA yields was assessed by a titration experiment using bone powder ranging from 50 to 1000mg. We obtained optimal DNA yields (27pgmg(-1) on average) when 150-200mg of starting material were processed using a one-step demineralisation method. Better-quality profiles (determined by the number of genotyped loci) were obtained when DNA was extracted from bone slices compared to extraction from bone powder. From bone slices 83.9% and from bone powder 46.7% of the samples provided genotypes for 11 or more loci. Since bone preparation procedures were carried out at room temperature, the method developed in the present study might be an attractive alternative to the standard freeze-mill approach, being faster and more cost-efficient.Fil: Caputo, Mariela. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina;Fil: Irisarri, Maximiliano. Universidad de Buenos Aires. Facultad de Farmacia y Bioquimica. Servicio de Huellas Digitales Geneticas;Fil: Alechine, Evguenia. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina;Fil: Corach, Daniel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina;Elsevier2013-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/1992Caputo, Mariela; Irisarri, Maximiliano; Alechine, Evguenia; Corach, Daniel; A DNA extraction method of small quantities of bone for high-quality genotyping; Elsevier; Forensic Science International: Genetics; 7; 5; 9-2013; 488-4931872-4973enginfo:eu-repo/semantics/altIdentifier/doi/doi:10.1016/j.fsigen.2013.05.002info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S1872497313001129info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:01:03Zoai:ri.conicet.gov.ar:11336/1992instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 10:01:04.285CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A DNA extraction method of small quantities of bone for high-quality genotyping
title A DNA extraction method of small quantities of bone for high-quality genotyping
spellingShingle A DNA extraction method of small quantities of bone for high-quality genotyping
Caputo, Mariela
Exhumed-Bone
Dna Extraction
Expert Software
Str Profile
title_short A DNA extraction method of small quantities of bone for high-quality genotyping
title_full A DNA extraction method of small quantities of bone for high-quality genotyping
title_fullStr A DNA extraction method of small quantities of bone for high-quality genotyping
title_full_unstemmed A DNA extraction method of small quantities of bone for high-quality genotyping
title_sort A DNA extraction method of small quantities of bone for high-quality genotyping
dc.creator.none.fl_str_mv Caputo, Mariela
Irisarri, Maximiliano
Alechine, Evguenia
Corach, Daniel
author Caputo, Mariela
author_facet Caputo, Mariela
Irisarri, Maximiliano
Alechine, Evguenia
Corach, Daniel
author_role author
author2 Irisarri, Maximiliano
Alechine, Evguenia
Corach, Daniel
author2_role author
author
author
dc.subject.none.fl_str_mv Exhumed-Bone
Dna Extraction
Expert Software
Str Profile
topic Exhumed-Bone
Dna Extraction
Expert Software
Str Profile
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv DNA genotyping techniques have been used successfully in forensic science for almost three decades and represent the gold standard for individual identification. However, efficient protocols for obtaining DNA from exhumed bones suitable for genotyping are still scarce and most of them require a considerable amount of starting material, are time consuming and are inefficient for reducing inhibitor's effects. We sought to develop an optimised protocol for extracting DNA from bone samples obtained from exhumations. We tested two approaches for preparing bone samples: (a) fine powder and (b) thin slices of bone. The best ratio of bone amount to DNA yields was assessed by a titration experiment using bone powder ranging from 50 to 1000mg. We obtained optimal DNA yields (27pgmg(-1) on average) when 150-200mg of starting material were processed using a one-step demineralisation method. Better-quality profiles (determined by the number of genotyped loci) were obtained when DNA was extracted from bone slices compared to extraction from bone powder. From bone slices 83.9% and from bone powder 46.7% of the samples provided genotypes for 11 or more loci. Since bone preparation procedures were carried out at room temperature, the method developed in the present study might be an attractive alternative to the standard freeze-mill approach, being faster and more cost-efficient.
Fil: Caputo, Mariela. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina;
Fil: Irisarri, Maximiliano. Universidad de Buenos Aires. Facultad de Farmacia y Bioquimica. Servicio de Huellas Digitales Geneticas;
Fil: Alechine, Evguenia. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina;
Fil: Corach, Daniel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina;
description DNA genotyping techniques have been used successfully in forensic science for almost three decades and represent the gold standard for individual identification. However, efficient protocols for obtaining DNA from exhumed bones suitable for genotyping are still scarce and most of them require a considerable amount of starting material, are time consuming and are inefficient for reducing inhibitor's effects. We sought to develop an optimised protocol for extracting DNA from bone samples obtained from exhumations. We tested two approaches for preparing bone samples: (a) fine powder and (b) thin slices of bone. The best ratio of bone amount to DNA yields was assessed by a titration experiment using bone powder ranging from 50 to 1000mg. We obtained optimal DNA yields (27pgmg(-1) on average) when 150-200mg of starting material were processed using a one-step demineralisation method. Better-quality profiles (determined by the number of genotyped loci) were obtained when DNA was extracted from bone slices compared to extraction from bone powder. From bone slices 83.9% and from bone powder 46.7% of the samples provided genotypes for 11 or more loci. Since bone preparation procedures were carried out at room temperature, the method developed in the present study might be an attractive alternative to the standard freeze-mill approach, being faster and more cost-efficient.
publishDate 2013
dc.date.none.fl_str_mv 2013-09
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/1992
Caputo, Mariela; Irisarri, Maximiliano; Alechine, Evguenia; Corach, Daniel; A DNA extraction method of small quantities of bone for high-quality genotyping; Elsevier; Forensic Science International: Genetics; 7; 5; 9-2013; 488-493
1872-4973
url http://hdl.handle.net/11336/1992
identifier_str_mv Caputo, Mariela; Irisarri, Maximiliano; Alechine, Evguenia; Corach, Daniel; A DNA extraction method of small quantities of bone for high-quality genotyping; Elsevier; Forensic Science International: Genetics; 7; 5; 9-2013; 488-493
1872-4973
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/doi:10.1016/j.fsigen.2013.05.002
info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S1872497313001129
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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