Término solicitado: SALOMON, OSCAR D./(22)
1 Registro/s encontrado/s en CAT+NORMA
| Tipo de Docum.: | artículo |
| Título: | Optimization of DNA extraction from individual sand flies for PCR amplification |
| Autor: | Caligiuri, Lorena G.; Sandoval, Adolfo E.; Miranda, Jose C.; Pessoa, Felipe A.; Santini, María S.; Salomón, Oscar D.; Secundino, Nagila F. C.; McCarthy, Christina B. |
| Título Ser./Col.: | Methods and Protocols, 20(2) |
| Autor Inst. Ser./Col.: | Multidisciplinary Digital Publishing Institute. MDPI. Basilea. Suiza |
| Idioma: | eng |
| Datos de Edición: | s.l. MDPI. 2019. |
| Pág./Vol.: | 15p. |
| Descriptores: | ADN; PCR; Biotecnología; Calcio |
| Resumen: | Numerous protocols have been published for extracting DNA from phlebotomines. Nevertheless, their small size is generally an issue in terms of yield, efficiency, and purity, for large-scale individual sand fly DNA extractions when using traditional methods. Even though this can be circumvented with commercial kits, these are generally cost-prohibitive for developing countries. We encountered these limitations when analyzing field-collected Lutzomyia spp. by polymerase chain reaction (PCR) and, for this reason, we evaluated various modifications on a previously published protocol, the most significant of which was a different lysis buffer that contained Ca2+ (buffer TESCa). This ion protects proteinase K against autolysis, increases its thermal stability, and could have a regulatory function for its substrate-binding site. Individual sand fly DNA extraction success was confirmed by amplification reactions using internal control primers that amplify a fragment of the cacophony gene. To the best of our knowledge, this is the first time a lysis buffer containing Ca2+ has been reported for the extraction of DNA from sand flies. |
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https://www.mdpi.com/2409-9279/2/2/36 |
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