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

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Resumen

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. 

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Biografía del autor/a (VER)

Juan D. Carvajal-Agudelo, Universidad de Caldas

 

 

M. Paula Trujillo-Betancur, Universidad de Caldas

 

 

Daniela Velásquez-Guarín, Universidad de Caldas

 

 

Hector E. Ramírez-Chaves, Universidad de Caldas

 

 

Jorge E. Pérez-Cárdenas, Universidad de Caldas

 

 

Fredy A. Rivera-Páez, Universidad de Caldas

 

 

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