Titulo:
Preservación en campo y extracción de ADN en sangre de mamíferos silvestres: métodos y factores claves para estudios de biodiversidad
.
Sumario:
Los estudios sobre salud pública y biodiversidad de mamíferos silvestres incluyen un componente genético. Para las muestras de sangre, se debe tener condiciones óptimas de colección, ya que pueden afectar la preservación y la extracción del ADN. Este estudio evaluó el uso de métodos de preservación de ADN líquido y seco y métodos de extracción de ADN comerciales y no comerciales, en muestras de sangre, recolectadas en campo. Para ello, se recogieron 264 muestras de sangre totales de mamíferos salvajes. Se preservó un primer grupo de muestras en clorhidrato de guanidina (GuHCl) y se extrajo el ADN, utilizando seis kits comerciales: Bioline, Norgen, Invitrogen, Promega y Qiagen, además de dos protocolos no comerciales: fenol-cloroformo isoami... Ver más
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Juan D. Carvajal-Agudelo, M. Paula Trujillo-Betancur, Daniela Velásquez-Guarín, Hector E. Ramírez-Chaves, Jorge E. Pérez-Cárdenas, Fredy A. Rivera-Páez - 2021
Esta obra está bajo una licencia internacional Creative Commons Atribución-NoComercial 4.0.
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UNIVERSIDAD DE CIENCIAS APLICADAS Y AMBIENTALES |
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Colombia |
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Revista U.D.C.A Actualidad & Divulgación Científica |
| title |
Preservación en campo y extracción de ADN en sangre de mamíferos silvestres: métodos y factores claves para estudios de biodiversidad |
| spellingShingle |
Preservación en campo y extracción de ADN en sangre de mamíferos silvestres: métodos y factores claves para estudios de biodiversidad Carvajal-Agudelo, Juan D. Trujillo-Betancur, M. Paula Velásquez-Guarín, Daniela Ramírez-Chaves, Hector E. Pérez-Cárdenas, Jorge E. Rivera-Páez, Fredy A. Biodiversidad Calidad de ADN Mammalia Sangre total Vida silvestre Biodiversity DNA quality Mammalia Whole blood Wildlife |
| title_short |
Preservación en campo y extracción de ADN en sangre de mamíferos silvestres: métodos y factores claves para estudios de biodiversidad |
| title_full |
Preservación en campo y extracción de ADN en sangre de mamíferos silvestres: métodos y factores claves para estudios de biodiversidad |
| title_fullStr |
Preservación en campo y extracción de ADN en sangre de mamíferos silvestres: métodos y factores claves para estudios de biodiversidad |
| title_full_unstemmed |
Preservación en campo y extracción de ADN en sangre de mamíferos silvestres: métodos y factores claves para estudios de biodiversidad |
| title_sort |
preservación en campo y extracción de adn en sangre de mamíferos silvestres: métodos y factores claves para estudios de biodiversidad |
| title_eng |
Field blood preservation and DNA extraction from wild mammals: methods and key factors for biodiversity studies |
| description |
Los estudios sobre salud pública y biodiversidad de mamíferos silvestres incluyen un componente genético. Para las muestras de sangre, se debe tener condiciones óptimas de colección, ya que pueden afectar la preservación y la extracción del ADN. Este estudio evaluó el uso de métodos de preservación de ADN líquido y seco y métodos de extracción de ADN comerciales y no comerciales, en muestras de sangre, recolectadas en campo. Para ello, se recogieron 264 muestras de sangre totales de mamíferos salvajes. Se preservó un primer grupo de muestras en clorhidrato de guanidina (GuHCl) y se extrajo el ADN, utilizando seis kits comerciales: Bioline, Norgen, Invitrogen, Promega y Qiagen, además de dos protocolos no comerciales: fenol-cloroformo isoamil alcohol (PC) y guanidina tiocianato (GIT). Otro grupo de muestras, se preservó en tarjetas Whatman® FTA® y se extrajo el ADN, con PC y GIT. Las extracciones con GIT y PC mostraron los valores y variaciones más altas en la concentración de ADN (ng/µL), mientras que el kit comercial mostró una baja variación. La preservación de la muestra en tarjetas Whatman® FTA® proporcionó una baja variación y cantidad de ADN extraído, en comparación con el uso de GuHCl. En cuanto a la calidad del ADN, los kits comerciales produjeron una mayor pureza (A260/280), mientras que los protocolos basados en GIT y PC proporcionaron resultados muy variables. Además, el uso de GIT y PC originó una mayor cantidad de ADN, pero de calidad variable. En general, la extracción basada en kits comerciales y la conservación Whatman® FTA® permitió obtener calidades y cantidades de ADN más estandarizadas. 
|
| description_eng |
Studies on public health and wild mammal biodiversity include a genetic component. For blood samples, there must be optimal sample collection conditions since these can affect DNA preservation and extraction. This study evaluated the use of liquid and dry DNA preservation methods and commercial and non-commercial DNA extraction methods on field-collected blood samples. For this, 264 total blood samples were collected from wild mammals. A first group of samples was preserved in guanidine hydrochloride (GuHCl) and DNA was extracted using six commercial kits:  Bioline, Norgen, Invitrogen, Promega, and Qiagen, in addition to phenol-chloroform isoamyl alcohol (PC) and guanidine thiocyanate (GIT). Another group of samples was preserved in Whatman® FTA® cards and DNA was extracted with PC and GIT. The extractions with GIT and PC showed the highest values (ng/µL) and variation in DNA concentration, while the commercial kit showed low variation. Sample preservation in Whatman® FTA® cards provided low variation and quantity of the extracted DNA compared with the use of GuHCl. Concerning DNA quality, the commercial kits yielded higher purity, while GIT and PC-based protocols provided highly variable results. Furthermore, the use of GIT and PC yielded a higher amount of DNA, yet, of variable quality. Overall, extraction based on commercial kits and Whatman® FTA® preservation allowed obtaining more standardized DNA qualities and quantities.
|
| author |
Carvajal-Agudelo, Juan D. Trujillo-Betancur, M. Paula Velásquez-Guarín, Daniela Ramírez-Chaves, Hector E. Pérez-Cárdenas, Jorge E. Rivera-Páez, Fredy A. |
| author_facet |
Carvajal-Agudelo, Juan D. Trujillo-Betancur, M. Paula Velásquez-Guarín, Daniela Ramírez-Chaves, Hector E. Pérez-Cárdenas, Jorge E. Rivera-Páez, Fredy A. |
| topicspa_str_mv |
Biodiversidad Calidad de ADN Mammalia Sangre total Vida silvestre |
| topic |
Biodiversidad Calidad de ADN Mammalia Sangre total Vida silvestre Biodiversity DNA quality Mammalia Whole blood Wildlife |
| topic_facet |
Biodiversidad Calidad de ADN Mammalia Sangre total Vida silvestre Biodiversity DNA quality Mammalia Whole blood Wildlife |
| citationvolume |
24 |
| citationissue |
1 |
| citationedition |
Núm. 1 , Año 2021 :Revista U.D.C.A Actualidad & Divulgación Científica. Enero-Junio |
| publisher |
Universidad de Ciencias Aplicadas y Ambientales U.D.C.A |
| ispartofjournal |
Revista U.D.C.A Actualidad & Divulgación Científica |
| source |
https://revistas.udca.edu.co/index.php/ruadc/article/view/1766 |
| language |
eng |
| format |
Article |
| rights |
http://creativecommons.org/licenses/by-nc/4.0 Juan D. Carvajal-Agudelo, M. Paula Trujillo-Betancur, Daniela Velásquez-Guarín, Hector E. Ramírez-Chaves, Jorge E. Pérez-Cárdenas, Fredy A. Rivera-Páez - 2021 Esta obra está bajo una licencia internacional Creative Commons Atribución-NoComercial 4.0. info:eu-repo/semantics/openAccess http://purl.org/coar/access_right/c_abf2 |
| references_eng |
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DNA extraction from dry museum beetles without conferring external morphological damage. PloS One. 2(3):e272. https://doi.org/10.1371/journal.pone.0000272 22. HAWKEY, C.M. 2017. Comparative mammalian haematology: cellular components and blood coagulation of captive wild animals. William Heinemann Medical Books. London, UK. 310p. https://doi.org/10.1016/C2013-0-06344-X 23. HENRY, P.; RUSSELLO, M.A. 2011. Obtaining high-quality DNA from elusive small mammals using low-tech hair snares. European J. Wildlife Research. 57(3):429-435. https://doi.org/10.1007/s10344-010-0449-y 24. HOFREITER, M. 2012. Nondestructive DNA extraction from museum specimens. Ancient DNA Springer. p.93-100. https://doi.org/10.1007/978-1-61779-516-9_13 25. IBRAHIM, N.A.; NASSAR, S.A.; ABD EL-GAWAD, A.M.; OMAR, M.F. 2018. Comparing the efficiency in DNA extraction between organic phenol and magnetic beads methods. Forensic Med. Toxicol. 16:10-17. https://doi.org/10.21608/zjfm.2018.2419.1007 26. KARTHIKEYAN, K.; SARANYA, R.; BHARATH, R.; VIDYA, R.; ITAMI, T.; SUDHAKARAN, R. 2020. A simple filter paper-based method for transporting and storing Enterocytozoon hepatopenaei DNA from infected Litopenaeus vannamei tissues. J. Invertebrate Pathology. 169:107305. https://doi.org/10.1016/j.jip.2019.107305 27. KRAVCHENKO, A.V.; CHETVERINA, E.V.; CHETVERIN, A.B. 2006. Preservation of nucleic acid integrity in guanidine thiocyanate lysates of whole blood. Russian J. Bioorganic Chemistry. 32(6):547-551. https://doi.org/10.1134/S1068162006060070 28. MA, D.; ZHUO, X.Y.; BU, J.; XIANG, P.; SHEN, B.H. 2007. Research of on the stability of ethanol in preservation of ethanol in blood. Fa Yi Xue Za Zhi. 23(2):117-119. 29. MALFERRARI, G.; MONFERINI, E.; DEBLASIO, P.; DIAFERIA, G.; SALTINI, G.; DEL VECCHIO, E.; ROSSI-BERNARDI, L.; BIUNNO, I. 2002. High-quality genomic DNA from human whole blood and mononuclear cells. Biotechniques. 33(6):1228-1230. https://doi.org/10.2144/02336bm09 30. 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Infectious diseases of wild mammals. John Wiley & Sons (Iowa, United States). 560p. https://doi.org/10.1002/9780470344880 |
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Preservación en campo y extracción de ADN en sangre de mamíferos silvestres: métodos y factores claves para estudios de biodiversidad Field blood preservation and DNA extraction from wild mammals: methods and key factors for biodiversity studies Los estudios sobre salud pública y biodiversidad de mamíferos silvestres incluyen un componente genético. Para las muestras de sangre, se debe tener condiciones óptimas de colección, ya que pueden afectar la preservación y la extracción del ADN. Este estudio evaluó el uso de métodos de preservación de ADN líquido y seco y métodos de extracción de ADN comerciales y no comerciales, en muestras de sangre, recolectadas en campo. Para ello, se recogieron 264 muestras de sangre totales de mamíferos salvajes. Se preservó un primer grupo de muestras en clorhidrato de guanidina (GuHCl) y se extrajo el ADN, utilizando seis kits comerciales: Bioline, Norgen, Invitrogen, Promega y Qiagen, además de dos protocolos no comerciales: fenol-cloroformo isoamil alcohol (PC) y guanidina tiocianato (GIT). Otro grupo de muestras, se preservó en tarjetas Whatman® FTA® y se extrajo el ADN, con PC y GIT. Las extracciones con GIT y PC mostraron los valores y variaciones más altas en la concentración de ADN (ng/µL), mientras que el kit comercial mostró una baja variación. La preservación de la muestra en tarjetas Whatman® FTA® proporcionó una baja variación y cantidad de ADN extraído, en comparación con el uso de GuHCl. En cuanto a la calidad del ADN, los kits comerciales produjeron una mayor pureza (A260/280), mientras que los protocolos basados en GIT y PC proporcionaron resultados muy variables. Además, el uso de GIT y PC originó una mayor cantidad de ADN, pero de calidad variable. En general, la extracción basada en kits comerciales y la conservación Whatman® FTA® permitió obtener calidades y cantidades de ADN más estandarizadas.  Studies on public health and wild mammal biodiversity include a genetic component. For blood samples, there must be optimal sample collection conditions since these can affect DNA preservation and extraction. This study evaluated the use of liquid and dry DNA preservation methods and commercial and non-commercial DNA extraction methods on field-collected blood samples. For this, 264 total blood samples were collected from wild mammals. A first group of samples was preserved in guanidine hydrochloride (GuHCl) and DNA was extracted using six commercial kits:  Bioline, Norgen, Invitrogen, Promega, and Qiagen, in addition to phenol-chloroform isoamyl alcohol (PC) and guanidine thiocyanate (GIT). Another group of samples was preserved in Whatman® FTA® cards and DNA was extracted with PC and GIT. The extractions with GIT and PC showed the highest values (ng/µL) and variation in DNA concentration, while the commercial kit showed low variation. Sample preservation in Whatman® FTA® cards provided low variation and quantity of the extracted DNA compared with the use of GuHCl. Concerning DNA quality, the commercial kits yielded higher purity, while GIT and PC-based protocols provided highly variable results. Furthermore, the use of GIT and PC yielded a higher amount of DNA, yet, of variable quality. Overall, extraction based on commercial kits and Whatman® FTA® preservation allowed obtaining more standardized DNA qualities and quantities. Carvajal-Agudelo, Juan D. Trujillo-Betancur, M. Paula Velásquez-Guarín, Daniela Ramírez-Chaves, Hector E. Pérez-Cárdenas, Jorge E. Rivera-Páez, Fredy A. Biodiversidad Calidad de ADN Mammalia Sangre total Vida silvestre Biodiversity DNA quality Mammalia Whole blood Wildlife 24 1 Núm. 1 , Año 2021 :Revista U.D.C.A Actualidad & Divulgación Científica. Enero-Junio Artículo de revista Journal article 2021-06-30T00:00:00Z 2021-06-30T00:00:00Z 2021-06-30 application/xml application/pdf Universidad de Ciencias Aplicadas y Ambientales U.D.C.A Revista U.D.C.A Actualidad & Divulgación Científica 0123-4226 2619-2551 https://revistas.udca.edu.co/index.php/ruadc/article/view/1766 10.31910/rudca.v24.n1.2021.1766 https://doi.org/10.31910/rudca.v24.n1.2021.1766 eng http://creativecommons.org/licenses/by-nc/4.0 Juan D. Carvajal-Agudelo, M. Paula Trujillo-Betancur, Daniela Velásquez-Guarín, Hector E. Ramírez-Chaves, Jorge E. Pérez-Cárdenas, Fredy A. Rivera-Páez - 2021 Esta obra está bajo una licencia internacional Creative Commons Atribución-NoComercial 4.0. AL-SHUHAIB, M.B.S.A. 2017. 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Infectious diseases of wild mammals. John Wiley & Sons (Iowa, United States). 560p. https://doi.org/10.1002/9780470344880 https://revistas.udca.edu.co/index.php/ruadc/article/download/1766/2134 https://revistas.udca.edu.co/index.php/ruadc/article/download/1766/2135 info:eu-repo/semantics/article http://purl.org/coar/resource_type/c_6501 http://purl.org/coar/resource_type/c_1843 info:eu-repo/semantics/publishedVersion http://purl.org/coar/version/c_970fb48d4fbd8a85 info:eu-repo/semantics/openAccess http://purl.org/coar/access_right/c_abf2 Text Publication |