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High quality DNA isolation method for chickpea genotypes

In chickpea breeding genetic studies of individual plants need to be evaluated at the DNA level using molecular markers. A simple and reliable DNA extraction method is a prerequisite. This small-scale method is cetyltrimethylammonium bromide (CTAB)-based and extracts DNA from 1 to 3 folded young leaves processed in a 1.5 ml tube with 0.5 ml of extraction buffer and homogenized using an electric drill. | Turk J Biol 29 (2005) 1-5 © TÜB‹TAK High Quality DNA Isolation Method for Chickpea Genotypes Hasibe C‹NG‹LL‹, Abdülkadir AKÇ‹N Gebze Institute of Technology, Department of Biology, 41400, Çay›rova, Gebze, Kocaeli - TURKEY Received: 17.03.2004 Abstract: In chickpea breeding genetic studies of individual plants need to be evaluated at the DNA level using molecular markers. A simple and reliable DNA extraction method is a prerequisite. This small-scale method is cetyltrimethylammonium bromide (CTAB)based and extracts DNA from 1 to 3 folded young leaves processed in a 1.5 ml tube with 0.5 ml of extraction buffer and homogenized using an electric drill. Compared with the micro-prep method the improved mini-prep CTAB method is highly efficient and much cheaper in terms of time, chemical use and labor input. About 49 samples per day can easily be processed by one person. The DNA yield is greater (60 µg per 50-100 µg of fresh leaf tissue) than that obtained from the micro-prep method (50 mg from 5 g of fresh leaf tissue). High quality DNA was obtained and used successfully for restriction endonuclease digestion and polymerase chain reaction amplifications using the mini-prep CTAB method. Key Words: Chickpea, DNA isolation, Molecular markers Abbreviations: CTAB, Cetyltrimethylammonium bromide; ISSR, Inter simple sequence repeat; PCR, Polymerase chain reaction; RAPD, Random amplified polymorphic DNA; SSR, Simple sequence repeat. Nohut Genotiplerinde Yüksek Kalitede Saf DNA ‹zolasyon Metodu Özet: Moleküler mark›rlardan faydalan›larak ekonomik aç›dan önemli birçok bitkinin ›slah edilmesi, günümüzde giderek önemi artan mark›r teknolojisinin ve mark›r yard›m›yla seleksiyon uygulamalar›n›n, art›k rutin çal›flmalar olabilece¤i konusunu gündeme getirmifltir. Bu nedenle, öncelikli olarak çal›flmalarda kullan›lacak ana materyal olan DNA’n›n, k›sa sürede ve saf olarak eldesi son derece önemlidir. Bunun için de güvenilir bir DNA izolasyon metodunun belirlenmesi gerekmektedir. Bu .