Evaluation of Four Different Protocols for DNA Extraction from Young and Old Carob Leaves

Abstract

Carob (Ceratonia siliqua L. (Cesalpinaceae) is an evergreen tree. It naturally grows in forests, but it is also commercially grown in some regions. As it was in other plant species, molecular markers are used to identify genetic diversity in carob. DNA isolation is the most significant factor effecting molecular marker analysis. In this study, four different isolation protocols were used to isolate quality DNA from young and old carob leaves. In young leaves, while the lowest DNA concentrations were obtained from protocol A (5.73±0.89 ng/µl DNA), DNA concentration of Protocol D were significantly higher than other protocols (900.66±23.38 ng/µl DNA). In old leaves, the lowest DNA concentrations were obtained from protocol A (2.9±0.4 ng/µl DNA) and the highest DNA concentrations were obtained from protocol C (11.36±0.8  ng/µl DNA). The DNA concentrations obtained from the freshest tip leaves measured 656.66±36.95 ng/µl DNA. DNA concentration of old leaves significantly lower than young leaves at all protocols. While the lowest A260/280 nm absorbance ratios in young leaves were observed in protocol A (0.76±0.02), the ratio in other protocols were between the optimum values. 

Keywords

• Carob,genetic diversity,DNA,molecular marker

Keçiboynuzunun Genç ve Olgun Yapraklarından DNA Ekstraksiyonu İçin Dört Farklı Protokolün Değerlendirilmesi

Öz

Keçiboynuzu (Ceratonia siliqua L. (Cesalpinaceae) herdem yeşil bir bitkidir. Keçiboynuzu doğal olarak ormanda yetişirken bazı bölgelerde ticari olarak da yetiştirilmektedir. Diğer bitki türlerinde olduğu gibi moleküler markırlar keçiboynuzunda da genetic çeşitliliği tanımlamak için kullanılmaktadır. Bu çalışmada, keçiboynozunun genç ve olgun yapraklarında dört farklı protocol kullanılarak kaliteli DNA elde etmek amaçlanmıştır.   Genç yapraklarda, En düşük DNA konsantrasyonu protocol A’dan (5.73±0.89 ng/µl DNA) sağlanırken, protocol D’nin (900.66±23.38 ng/µl DNA) DNA konsantrasyonu diğer protokollerden oldukça yüksektir. Yaşlı yapraklarda, en düşük DNA miktarı protokol A(2.9±0.4 ng/µl DNA) ile en yüksek DNA miktarı protokol C(11.36±0.8  ng/µl DNA) şeklindedir. Genç yapraklardan elde edilen DNA konsantrasyonu 656.66±36.95 ng/µl DNA olarak ölçülmüştür. Olgun yapraklardaki DNA konsantrasyonları genç yapraklardaki bütün protokollere göre oldukça düşüktür. Genç yapraklarda en düşük emilim A260/280 nm ile protokol A (0.76±0.02) iken, diğer protokollerdeki oranlar optimum değerler arasındadır.  

Anahtar Kelimeler

• Keçiboynuzu,genetic çeşitlilik,DNA,moleküler markır

Kaynakça

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