Amélioration de la méthode d'extraction d'ADN au CTAB appliquée aux feuilles de cotonnier

• Main Authors: Mergeai G., Baudoin JP., Benbouza H.
• Format: Article
• Language: English
• Published: Presses Agronomiques de Gembloux 2006-01-01
• Series: Biotechnologie, Agronomie, Société et Environnement
• Subjects: DNA extraction; Gossypium hirsutum L.; CTAB; microsatellites; activated charcoal; recalcitrant
• Online Access: http://www.pressesagro.be/base/text/v10n2/73.pdf
• ISSN: 1370-6233; 1780-4507

Improvement of the genomic DNA extraction method with CTAB for cotton leaves. Molecular genetic analysis in cotton (Gossypium hirsutum L.) is often limited by the availability of fresh tissue and the time necessary to extract DNA from it. To overcome these problems, the original CTAB method was improved. The major modifi cations concern DNA precipitation at -20°C, incubation of the resuspended DNA at 60°C and centrifugation at 4°C for all extraction steps. The improved method was relatively fast, cheap and yielded high quality DNA (80-200 μg . g-1). The optimized protocol gives satisfactory results on dried and frozen leaves at -80°C. The DNA was suitable for restriction-enzyme digestion with EcoRI and as a template for polymerase chain reaction (PCR) using more than two hundred microsatellite cotton primers on different cotton species, the G. hirsutum × G. raimondii × G. sturtianum trispecifi c hybrid and its progenies.