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

Published online by Cambridge University Press:  28 February 2007

José Rivero ,
Ludmel Urdaneta ,
Normig Zoghbi ,
Martha Pernalete ,
Yasmin Rubio-Palis  and
Flor Herrera
José Rivero
Affiliation:
Facultad de Ciencias de la Salud, Centro de Investigaciones Biomédicas BIOMED, Universidad de Carabobo, Núcleo Aragua, Venezuela
Ludmel Urdaneta
Affiliation:
Facultad de Ciencias de la Salud, Centro de Investigaciones Biomédicas BIOMED, Universidad de Carabobo, Núcleo Aragua, Venezuela
Normig Zoghbi
Affiliation:
Facultad de Ciencias de la Salud, Centro de Investigaciones Biomédicas BIOMED, Universidad de Carabobo, Núcleo Aragua, Venezuela
Martha Pernalete
Affiliation:
Facultad de Ciencias de la Salud, Centro de Investigaciones Biomédicas BIOMED, Universidad de Carabobo, Núcleo Aragua, Venezuela
Yasmin Rubio-Palis
Affiliation:
Facultad de Ciencias de la Salud, Centro de Investigaciones Biomédicas BIOMED, Universidad de Carabobo, Núcleo Aragua, Venezuela Instituto de Altos Estudios ‘Dr Arnoldo Gabaldón’, Ministerio de Salud y Desarrollo Social (Ministry of Health and Social Development), Maracay, Aragua, Venezuela
Flor Herrera*
Affiliation:
Facultad de Ciencias de la Salud, Centro de Investigaciones Biomédicas BIOMED, Universidad de Carabobo, Núcleo Aragua, Venezuela
*
*Email: [email protected]
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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.

Keywords

Anopheles darlingiAedes aegyptiDNA integrityDNA extractionmosquito DNA
Type
Short Communication
Information
International Journal of Tropical Insect Science , Volume 24 , Issue 3 , September 2004 , pp. 266 - 269
Copyright
Copyright © ICIPE 2004

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References

Ballinger-Crabtree, M. E., Black, I. V. W. C., Miller, B. R. (1992) Use of genetic polymorphisms detected by the random-amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) for differentiation and identification of Aedes aegypti subspecies and populations. Am. J. Trop. Med. Hyg. 47, 893–901.CrossRefGoogle ScholarPubMed
Black, W. C., Munstermann, L. M. (1996) Molecular taxonomy and systematics of arthropod vectors, 438–470. In Biology of Disease Vectors. (Edited by Marquardt, W. C., Beaty, B.). University Press of Colorado, Boulder, Colorado.Google Scholar
Coen, E. S., Strachan, T., Dover, G. (1982) Dynamics of concerted evolution of ribosomal DNA and histone gene families in the melanogaster species subgroup of Drosophila. J. Mol. Biol. 15, 17–35.Google Scholar
Manguin, S., Wilkerson, R., Conn, J., Rubio-Palis, Y., Danoff-Burg, J., Roberts, D. (1999) Population structure of the primary malaria vector in South America, Anopheles darlingi, using isozyme, random amplified polymorphic DNA, internal transcribed spacer 2, and morphological markers. Am. J. Trop. Med. Hyg. 60, 364–376.CrossRefGoogle Scholar
Nadeau, J. H., Bedigian, H. G., Bouchard, G., Denial, T., Kosowsky, M., Norberg, R., Pugh, S., Sargeant, E., Turner, R., Paigen, B. (1992) Multilocus markers for mouse genome analysis: PCR amplification based on single primers of arbitrary nucleotide sequence. Mamm. Genome 3, 55–64.Google Scholar
Posso, C. E., Gonzalez, R., Cardenas, H., Gallego, G., Duque, M. C., Suarez, M. F. (2003) Random amplified polymorphic DNA analysis of Anopheles nuneztovari (Diptera: Culicidae) from western and northeastern Colombia. Mem. Inst. Oswaldo Cruz 98, 469–476.CrossRefGoogle ScholarPubMed