Técnica de extracción de adn a partir de la glándula de seda del gusano de seda bombyx morí (lepidoptera: bombycidae)
Mejía Cruz, Rodolfo Alberto | 2006-01
Selection for production of puré lines of the silkworm (Bombyx morí), starting with commercial hybrids, is a long and expensive process, however, molecular biology techniques can shorten the time and allow to determine favorable characteristics for silk production. The objective of this work consisted in establishing a protocol for the extraction of DNA of good quality and quantity of the silkworm. The research was carried out at the molecular biology laboratory of the Ü.D.C.A at Bogotá, using larvae coming from the breeds of segregant lines maintained in the laboratory. For the extraction of DMA the posterior, médium and anterior part, placed in a Buffer extraction solution with Tris HCI, MaCI and EDTA pH 8, of the silk gland was used. Subsequently the samples were washed with phenol chloroform and isoamylic chloroform. For the DMA precipitation frozen ethanol was applied. Electrophoresis in agarosa was performed and quantified in a mini-fluorometer with a fixed longitude wave, obtaining a range between 0.21/xg/ml and 1.5/xg/ml of DMA. The quality and quantity of the DMA of the posterior part of the silk gland of five day oíd larvae of the last instar was superior to that obtained from larvae of other ages and other segments of the gland. Therefore the posterior part of the silk gland is considered to be the most adequate tissue for the DMA extraction process related to molecular analysis of silk productivity components.