DNA extraction methods from vicuña fecal samples (Vicugna vicugna mensalis)

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At present, the use of DNA extracted from non-invasive samples, especially feces, is common practice in the discipline of conservation genetics. Even though DNA analysis is often difficult mainly due to the composition and preservation of feces has been shown in many species with the development of...

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Detalles Bibliográficos
Autores: Maturrano H., Lenin, Aguilar L., Juan M., Krüger D., Paloma, Chávez I., Jesús, Rosadio A., Raúl, Wheeler, Jane C.
Formato: artículo
Fecha de Publicación:2012
Institución:Universidad Nacional Mayor de San Marcos
Repositorio:Revistas - Universidad Nacional Mayor de San Marcos
Lenguaje:español
OAI Identifier:oai:ojs.csi.unmsm:article/918
Enlace del recurso:https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/918
Nivel de acceso:acceso abierto
Materia:fecal DNA extraction
feces
vicuña
Extracción de ADN
heces
vicuña.
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oai_identifier_str oai:ojs.csi.unmsm:article/918
network_acronym_str REVUNMSM
network_name_str Revistas - Universidad Nacional Mayor de San Marcos
repository_id_str
dc.title.none.fl_str_mv DNA extraction methods from vicuña fecal samples (Vicugna vicugna mensalis)
OPTIMIZACIÓN DE UNA TÉCNICA PARA LA EXTRACCIÓN DE UN ADN DE HECES DE VICUÑA (Vicugna vicugna mensalis).
title DNA extraction methods from vicuña fecal samples (Vicugna vicugna mensalis)
spellingShingle DNA extraction methods from vicuña fecal samples (Vicugna vicugna mensalis)
Maturrano H., Lenin
fecal DNA extraction
feces
vicuña
Extracción de ADN
heces
vicuña.
title_short DNA extraction methods from vicuña fecal samples (Vicugna vicugna mensalis)
title_full DNA extraction methods from vicuña fecal samples (Vicugna vicugna mensalis)
title_fullStr DNA extraction methods from vicuña fecal samples (Vicugna vicugna mensalis)
title_full_unstemmed DNA extraction methods from vicuña fecal samples (Vicugna vicugna mensalis)
title_sort DNA extraction methods from vicuña fecal samples (Vicugna vicugna mensalis)
dc.creator.none.fl_str_mv Maturrano H., Lenin
Aguilar L., Juan M.
Krüger D., Paloma
Chávez I., Jesús
Rosadio A., Raúl
Wheeler, Jane C.
author Maturrano H., Lenin
author_facet Maturrano H., Lenin
Aguilar L., Juan M.
Krüger D., Paloma
Chávez I., Jesús
Rosadio A., Raúl
Wheeler, Jane C.
author_role author
author2 Aguilar L., Juan M.
Krüger D., Paloma
Chávez I., Jesús
Rosadio A., Raúl
Wheeler, Jane C.
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv fecal DNA extraction
feces
vicuña
Extracción de ADN
heces
vicuña.
topic fecal DNA extraction
feces
vicuña
Extracción de ADN
heces
vicuña.
description At present, the use of DNA extracted from non-invasive samples, especially feces, is common practice in the discipline of conservation genetics. Even though DNA analysis is often difficult mainly due to the composition and preservation of feces has been shown in many species with the development of suitable extraction methods is possible to maximize the reliability and effectiveness of such procedures. The objective of this study has been optimization of a protocol for the extraction of DNA from vicuña feces (Vicugna vicugna mensalis), species classified as Near Threatened in Peru, in order to obtain DNA of sufficient quality for molecular analysis. A total of 51 samples were taken from vicuña dung piles in two communities located 4600 meters above sea level in the dry Puna ecosystems of the regions of Junin and Moquegua between August and September 2008. After observing defecation, samples were collected immediately, preserved in ethanol and transported to the laboratory for DNA extraction. Modification of the protocol of the QIAamp ® DNA Stool Mini Kit (QIAGEN) to include an extended period of vigorous shaking was found to producer higher quality DNA than was obtained using both the kit protocol and PCI (Phenol/chloroform/isoamyl alcohol). The amount of extracted DNA was visualized using horizontal agarose gel electrophoresis, and the quality was tested analyzing microsatellite markers LCA 19 and YWLL 46 on polyacrylamide gels. Although DNA was obtained by all three methods, only the modified protocol produced DNA of sufficient quality for use in molecular analyses.
publishDate 2012
dc.date.none.fl_str_mv 2012-09-28
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/918
10.15381/rivep.v23i3.918
url https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/918
identifier_str_mv 10.15381/rivep.v23i3.918
dc.language.none.fl_str_mv spa
language spa
dc.relation.none.fl_str_mv https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/918/743
dc.rights.none.fl_str_mv https://creativecommons.org/licenses/by-nc-sa/4.0
info:eu-repo/semantics/openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/4.0
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidad Nacional Mayor de San Marcos, Facultad de Medicina Veterinaria
publisher.none.fl_str_mv Universidad Nacional Mayor de San Marcos, Facultad de Medicina Veterinaria
dc.source.none.fl_str_mv Revista de Investigaciones Veterinarias del Perú; Vol. 23 Núm. 3 (2012); 369-376
Revista de Investigaciones Veterinarias del Perú; Vol. 23 No. 3 (2012); 369-376
1682-3419
1609-9117
reponame:Revistas - Universidad Nacional Mayor de San Marcos
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instacron_str UNMSM
institution UNMSM
reponame_str Revistas - Universidad Nacional Mayor de San Marcos
collection Revistas - Universidad Nacional Mayor de San Marcos
repository.name.fl_str_mv
repository.mail.fl_str_mv
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spelling DNA extraction methods from vicuña fecal samples (Vicugna vicugna mensalis)OPTIMIZACIÓN DE UNA TÉCNICA PARA LA EXTRACCIÓN DE UN ADN DE HECES DE VICUÑA (Vicugna vicugna mensalis).Maturrano H., LeninAguilar L., Juan M.Krüger D., PalomaChávez I., JesúsRosadio A., RaúlWheeler, Jane C.fecal DNA extractionfecesvicuñaExtracción de ADNhecesvicuña.At present, the use of DNA extracted from non-invasive samples, especially feces, is common practice in the discipline of conservation genetics. Even though DNA analysis is often difficult mainly due to the composition and preservation of feces has been shown in many species with the development of suitable extraction methods is possible to maximize the reliability and effectiveness of such procedures. The objective of this study has been optimization of a protocol for the extraction of DNA from vicuña feces (Vicugna vicugna mensalis), species classified as Near Threatened in Peru, in order to obtain DNA of sufficient quality for molecular analysis. A total of 51 samples were taken from vicuña dung piles in two communities located 4600 meters above sea level in the dry Puna ecosystems of the regions of Junin and Moquegua between August and September 2008. After observing defecation, samples were collected immediately, preserved in ethanol and transported to the laboratory for DNA extraction. Modification of the protocol of the QIAamp ® DNA Stool Mini Kit (QIAGEN) to include an extended period of vigorous shaking was found to producer higher quality DNA than was obtained using both the kit protocol and PCI (Phenol/chloroform/isoamyl alcohol). The amount of extracted DNA was visualized using horizontal agarose gel electrophoresis, and the quality was tested analyzing microsatellite markers LCA 19 and YWLL 46 on polyacrylamide gels. Although DNA was obtained by all three methods, only the modified protocol produced DNA of sufficient quality for use in molecular analyses.Actualmente es posible determinar el estatus de especies silvestres mediante el análisis de ADN extraído de muestras no invasivas como las heces. Aunque dichos análisis suelen ser dificultosos debido principalmente a la composición y conservación de las heces, se ha demostrado en muchas especies que con el desarrollo de métodos adecuados de extracción es posible maximizar la fiabilidad y eficacia de este procedimiento. Por tanto, el objetivo del presente estudio fue optimizar un método de extracción de ADN de heces de vicuña Vicugna vicugna mensalis, especie clasificada como casi amenazada en el Perú, con la finalidad de obtener ADN de suficiente calidad para análisis moleculares. Se recolectaron 51 muestras de heces durante los meses de agosto y septiembre de 2008 en dos comunidades vicuñeras ubicadas a más de 4600 msnm en los departamentos de Junín y Moquegua. Luego de observar la defecación, las muestras fueron recolectadas en forma inmediata, conservadas en etanol y transportadas al laboratorio para la extracción del ADN. El kit comercial QIAamp® DNA Stool Mini Kit (QIAGEN) fue modificado para incluir un vigoroso y extenso proceso de agitación y los resultados fueron comparados con dos protocolos comúnmente utilizados: fenol/cloroformo/alcohol isoamílico y el mismo kit QIAGEN. La cantidad del ADN extraído fue observado en electroforesis horizontal en geles de agarosa, y la calidad evaluada mediante la amplificación y visualización de los tamaños esperados de los microsatélites YWLL46 y LCA19. Aunque se logró extraer ADN con los tres métodos, solamente el método modificado permitió obtener ADN de suficiente calidad para ser utilizado en pruebas moleculares.Universidad Nacional Mayor de San Marcos, Facultad de Medicina Veterinaria2012-09-28info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/91810.15381/rivep.v23i3.918Revista de Investigaciones Veterinarias del Perú; Vol. 23 Núm. 3 (2012); 369-376Revista de Investigaciones Veterinarias del Perú; Vol. 23 No. 3 (2012); 369-3761682-34191609-9117reponame:Revistas - Universidad Nacional Mayor de San Marcosinstname:Universidad Nacional Mayor de San Marcosinstacron:UNMSMspahttps://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/918/743Derechos de autor 2012 Lenin Maturrano H., Juan M. Aguilar L., Paloma Krüger D., Jesús Chávez I., Raúl Rosadio A., Jane C. Wheelerhttps://creativecommons.org/licenses/by-nc-sa/4.0info:eu-repo/semantics/openAccessoai:ojs.csi.unmsm:article/9182020-03-19T19:34:52Z
score 13.914502
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