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A Simple and Rapid Method for Mini Preparation of High Molecular Weight DNA from Certain Acarines, Bacteria and Soybean

Published online by Cambridge University Press:  19 September 2011

Z. A. El-Fiky
Affiliation:
Genetics Department, Faculty of Agriculture, Fayoum Branch Cairo University, Fayoum, Egypt
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Abstract

A simple, rapid and low-cost method for the isolation of high molecular weight DNA from small amounts of cells or tissue is described. The method does not require liquid nitrogen, proteinase K or costly buffers. This method was successfully applied in the isolation and purification of DNA from eggs of Hyalomma excavatum, H. dromedarii, Argas persicus, A. hermanii and Boophilus annulatus, in addition to Escherichia coli, actinomycetes (Nocardia sp.) and leaves of soybean (Glycine max L.). The DNA concentration was calculated from the optical density (OD) value at wavelength 260 nm and the ratios OD260/OD280 and OD260/OD280 determined. The quality and amount of DNA obtained by this method were comparable to those of DNA isolated from equal amounts of material using a commercial genomie DNA purification kit.

Résumé

Cette publication présente une méthode simple, rapide et peu coûteuse d'isolement de l'ADN de poids moléculaire élevé à partir d'une petite quantité de cellules ou de tissus. La méthode ne nécessite ni hydrogène liquide, ni protéinase K, ni même de tampons coûteux. Cette méthode a été appliquée avec succès pour l'isolement et la purification d'ADN d'oeufs de Hyalomma excavatum, de H. dromedarii, d'Argas persicus, d'A. hermanii et de Boophilus annulatus, ainsi que d'Escherichia coli, de l'actinomycète (Nocardia sp.) et de feuilles de soja (Glycine max L.). La concentration d'ADN est calculée par la méthode des densités optiques (DO) pour une longueur d'onde de 260 nm et les rapports DO260/DO280etDO260/DO230 sont déterminés. La qualité et la quantité d'ADN obtenues par cette méthode sont comparables à celles isolées de quantités identiques de matériel à partir d'un kit commercial de purification d'ADN génomique.

Type
Research Articles
Copyright
Copyright © ICIPE 2003

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