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International Journal of Tropical Insect Science

, Volume 24, Issue 3, pp 266–269 | Cite as

Optimization of extraction procedure for mosquito DNA suitable for PCR-based techniques

  • José Rivero
  • Ludmel Urdaneta
  • Normig Zoghbi
  • Martha Pernalete
  • Yasmin Rubio-Palis
  • Flor HerreraEmail author
Short Communication

Abstract

A modified technique to extract intact DNA from Aedes aegypti and Anopheles darlingi mosquitoes is presented. The samples were treated initially with proteinase K to digest all proteinaceous matter. For both vectors, the optimum temperature for the protease treatment was 55°C and the best pre-incubation time was 4 h. RNA contamination was eliminated with RNAse treatment. The purity of the DNA was high since the A260/A280 ratio averaged >1.80 for all samples and the quantity of DNA extracted was >1.6 times higher than that using the unmodified procedure. The DNA obtained displayed clear random amplified polymorphic DNA (RAPD) profiles, which indicates that it can be used for polymerase chain reaction (PCR)-based techniques.

Key words

Anopheles darlingi Aedes aegypti DNA integrity DNA extraction mosquito DNA 

Mots clés

Anopheles darlingi Aedes aegypti intégrité de l’ADN extraction de l’ADN ADN de moustique 

Résumé

Cette étude présente une méthode modifiée pour l’extraction de l’ADN de haute qualité à partir de moustiques de Aedes aegypti et Anopheles darlingi. Les souches étaient traitées initialement avec proteinase K pour digérer toutes les matières protéinaseus. Ce traitement protéique montrait une température optimale de 55°C et son meilleur temps d’incubation de 4 heures pour les deux vecteurs. La pureté de l’ADN était haute puisque la moyenne de la relation A260/A280 était >1.80 pour toutes les souches. L’ADN obtenu montrait de clairs profils de RAPD ce qui indique qu’il peut être utilisé pour les techniques de PCR.

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Copyright information

© ICIPE 2004

Authors and Affiliations

  • José Rivero
    • 1
  • Ludmel Urdaneta
    • 1
  • Normig Zoghbi
    • 1
  • Martha Pernalete
    • 1
  • Yasmin Rubio-Palis
    • 1
    • 2
  • Flor Herrera
    • 1
    Email author
  1. 1.Facultad de Ciencias de la Salud, Centro de Investigaciones Biomédicas BIOMEDUniversidad de CaraboboNúcleo AraguaVenezuela
  2. 2.Instituto de Altos Estudios ‘Dr Arnoldo Gabaldón’, Ministerio de Salud y Desarrollo Social (Ministry of Health and Social Development)Maracay AraguaVenezuela

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