Optimization of extraction procedure for mosquito DNA suitable for PCR-based techniques
Autor(es): Herrera, F.; Rivero, J.; Urdaneta, L.; Zoghbi, N.; Pernalete, M.; Rubio-Palis, Y.
Resumo: 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 degree 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 A sub(260)/A sub(280) 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.
Palavras-Chave: Mosquitoes; Yellow fever mosquito
Imprenta: International Journal of Tropical Insect Science, v. 24, n. 3, p. 266-269, 2004.
Descritores: Aedes aegypti - DNA ; Aedes aegypti - RNA
Data de publicação: 2004
