Developing a DNA isolation protocol specific to Pinus species distributed in Turkey

In this thesis, an optimized protocol was developed to achieve high-yield and high-quality DNA isolation from different Pinus species. The study involved five main Pinus species native to Turkey's natural habitats (Pinus brutia, Pinus nigra, Pinus sylvestris, Pinus halepensis, and Pinus pinea) and two non-native species (Pinus pinaster and Pinus radiata). The yield and quality of the isolated DNA samples were evaluated using spectrophotometric methods, and differences among species were analyzed using statistical methods. The Shapiro-Wilk normality test indicated that the data did not follow a normal distribution, leading to the use of non-parametric analysis methods. Results from the Kruskal-Wallis test and Dunn-Bonferroni post-hoc test revealed statistically significant differences among species in terms of DNA yield and quality. The integrity of the obtained DNA samples was assessed via agarose gel electrophoresis, resulting in high-molecular-weight, intact DNA bands. Additionally, PCR amplifications were successfully performed, and the amplicons were confirmed through gel electrophoresis. The isolated DNA samples were suitable for sequencing applications, and sequencing results were successfully obtained. These findings demonstrate that the optimized protocol can be effectively used in both traditional molecular techniques and advanced genetic analyses. The protocol offers significant advantages due to its short application time (approximately 65 minutes) and low cost (approximately $0.05 per sample). Compared to commercial kits, its cost-effectiveness and consistent results across different Pinus species enhance its desirability for large-scale genetic and molecular biology studies. Furthermore, it is recommended that future studies explore the applicability of the protocol to different plant species and tissues.