Comparación de los métodos de preservación de especímenes Anopheles (Diptera: Culicidae) para la extracción de ADN.

Introduction: Various methodologies have been reported for long term specimen preservation, which protect the DNA from degradation allowing its posterior analysis in molecular studies. However, the effectiveness of the preservation methods may vary among insect groups; therefore, it is convenient to...

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Publicado en:Salud Uninorte Vol. 27; no. 1; pp. 36 - 49
Autores principales: Sepúlveda, Tatiana Castaño, Zapata, Paola Sánchez, Guerra, Lorena Viana, Ochoa, Margarita Correa, Builes, Lina Gutiérrez, Tamayo, Mario Zapata
Formato: Artículo
Publicado: Fundacion Universidad del Norte ene-jun2011
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Acceso en línea:Ver este registro en EBSCOhost
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        au:
          Sepúlveda, Tatiana Castaño
          Zapata, Paola Sánchez
          Guerra, Lorena Viana
          Ochoa, Margarita Correa
          Builes, Lina Gutiérrez
          Tamayo, Mario Zapata
        affil:
          Grupo de Microbiología Molecular, Escuela de Microbiología, Universidad de Antioquia. Medellín (Colombia).
          Grupo de Investigación en Microbiología Veterinaria, Escuela de Microbiología, Universidad de Antioquia. Medellín (Colombia).
      su:
        Anopheles
        Mosquitoes
        Biological specimens
        Preservation of materials
        DNA
        Malaria
        Polymerase chain reaction
      sug:
        subj:
          Anopheles
          Mosquitoes
          Biological specimens
          Preservation of materials
          DNA
          Malaria
          Polymerase chain reaction
      keyword:
        malaria
        polymerase chain reaction
        samples preservation
      ab: Introduction: Various methodologies have been reported for long term specimen preservation, which protect the DNA from degradation allowing its posterior analysis in molecular studies. However, the effectiveness of the preservation methods may vary among insect groups; therefore, it is convenient to determine the preservation method to be used with a particular group before performing molecular studies with a large number of specimens. Objetives: An experimental comparative study was conducted to evaluate different preservation treatments for Anopheles albimanus, to know their utility in conserving the quantity and quality of DNA to be used in subsequent molecular studies. Methods: DNA was extracted from each specimen, it was then quantified by spectrophotometer and the Internal Transcribed Spacer 2 (ITS2) was amplified by PCR. Results: ITS2 amplification was obtained in all specimens without previous preservation, in 60% of those preserved in sílica gel and in 20% of those preserved at -80°C and in 70% ethanol. A significant difference (p<0.05) was found between amplification proportions. Amplification was not obtained in specimens preserved frozen at -20°C and not correlation by logistic regression was observed between the DO260/DO280 index and DNA concentration of specimens presenting ITS2 amplification. Conclusions: The data suggest that keeping specimens in silica gel or frozen at -80°C may be the best conditions to preserve Anopheles specimens for subsequent molecular studies.
      pubtype: Academic Journal
      doctype: Article
      src: R
    language: Spanish
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