Comparación de los métodos de preservación de especímenes Anopheles (Diptera: Culicidae) para la extracción de ADN
Journal Title: Revista Científica Salud Uninorte - Year 2011, Vol 27, Issue 1
Abstract
Abstract Introduction: Various methodologies have been reported for long term specimen preser- vation, which protect the DNA from degradation allowing its posterior analysis in molecu- lar 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 pres- ervation 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 spectropho- tometer 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 correla- tion by logistic regression was observed between the DO260/DO280 index and DNA con- centration 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. Key words: Anopheles, malaria, DNA, samples preservation, polymerase chain reaction.
Authors and Affiliations
Tatiana Castaño Seúlveda, Paola sánchez Zapata, Lorena Viana Guerra, Margarita Correa Ochoa, Lina Gutierrez Builes, Maria Zapata Tamayo
Keywords
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