Makarnalik Buğday Genotiplerinin Genetic Çeşitliliğinin Saptanması için iPBS Işaretleyicilerine Dayalı PCR Analizinin Optimizasyonu

Abstract

Bu çalışmada, makarnalık buğday genotiplerinin genetik çeşitliliğini belirlemek amacıyla iPBS (inter-primer binding site) moleküler belirteçleri kullanılarak PCR optimizasyonu gerçekleştirilmiştir. iPBS belirteçlerinin etkinliğini artırmak için PCR koşullarının optimize edilmesi kritik bir aşama olmuştur. Moleküler analizlerin doğruluğunu artırmak adına, 20 farklı iPBS primeriyle yapılan gradient testi ile optimum annealing sıcaklıkları belirlenmiştir. Elde edilen sonuçlara göre, 11 primerin polimorfik bantlar oluşturduğu ve bu primerlerin en verimli şekilde bağlandığı sıcaklıklar tespit edilmiştir. Primerlerin bağlanma sıcaklıklarının doğru bir şekilde belirlenmesi, PCR sürecinin başarısını doğrudan etkileyerek güvenilir DNA amplifikasyonu elde edilmesini sağlamıştır.

Keywords

• Makarnalık buğday
• moleküler belirteçler
• PCR optimizasyonu

PCR Optimization Based on Inter-Primer Binding Site (iPBS) Markers for Genetic Diversity Analysis in Durum Wheat Genotypes

Abstract

In this study, PCR optimization was performed using iPBS (inter-primer binding site) molecular markers to determine the genetic diversity of durum wheat genotypes. Optimizing PCR conditions to enhance the efficiency of iPBS markers was a critical step. To improve the accuracy of the molecular analysis, gradient testing was conducted with 20 different iPBS primers to determine the optimum annealing temperatures. The results showed that 11 primers produced polymorphic bands, and the temperatures at which these primers bound most efficiently were identified. Correctly determining the annealing temperatures of the primers directly impacted the success of the PCR process, ensuring reliable DNA amplification.

Keywords

• Durum wheat
• molecular markers
• PCR optimization

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