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

Year 2017, Volume: 48 Issue: 2, 73 - 79, 14.10.2017

Hasan Pinar Mehmet Yaman Hakan Keleş Mustafa Ünlü Ömer Faruk Coşkun

https://doi.org/10.17097/ataunizfd.304430

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

Abstract

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.  

Keywords

Keçiboynuzu, genetic çeşitlilik, DNA, moleküler markır

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