Ay denizanası Aurelia aurita'nın (Linnaeus, 1758) farklı dokularında DNA izolasyon yöntemlerinin verimliliklerinin değerlendirilmesi

Abstract

Özet Amaç: Bu çalışma, jelatinimsi yapısı nedeniyle DNA izolasyonu zor olan denizanası Aurelia aurita’dan elde edilecek DNA verimini optimize etmeyi; doku türü, enzimatik inkübasyon süresi ve farklı izolasyon yöntemlerinin etkilerini karşılaştırarak değerlendirmeyi amaçlamıştır. Yöntem: Toplamda üç farklı DNA izolasyon yöntemi (salting-out, kit bazlı ve Trizol bazlı) dört farklı doku türü (gonad, tentakül, epidermis ve gastrodermis) üzerinde uygulanmış, her biri Proteinaz K ile 0, 1, 3 ve 24 saat inkübasyona tabi tutulmuştur. Böylece 48 farklı kombinasyon analiz edilmiştir. Bulgular: En yüksek DNA konsantrasyonları gonad dokularında elde edilmiş olup bu durum, dokunun yüksek hücresel yoğunluğu ve yapısal bütünlüğü ile ilişkilendirilmiştir. Özellikle 1–3 saatlik Proteinaz K uygulamasıyla birlikte salting-out yöntemi, diğer iki yönteme kıyasla daha yüksek verim sağlamıştır. Aşırı nemli olan epidermis ve gastrodermis dokularında ise yaş ağırlık ölçümünde sapmalar oluşmuş ve bu durum biyolojik girdiye göre DNA veriminin olduğundan düşük hesaplanmasına neden olmuştur. Sonuç: Jelatinöz denizel omurgasızlardan DNA izolasyonunda doku seçimi, enzimatik inkübasyon süresi ve kullanılan metodoloji büyük önem taşımaktadır. Aurelia aurita örneklerinde, özellikle gonad dokusuna 1–3 saatlik Proteinaz K uygulaması ile salting-out yöntemi kullanımı, verimli ve güvenilir bir DNA izolasyonu için önerilmektedir.

Keywords

• DNA izolasyonu
• DNA ekstraksiyonu
• Aurelia aurita
• tuzla çöktürme yöntemi
• DNA verimi

Project Number

FLO-2024-40848

Evaluating yield of DNA extraction methods across different tissues of moon jellyfish Aurelia aurita (Linnaeus, 1758)

Abstract

Purpose: This study aimed to optimize DNA extraction from the moon jellyfish Aurelia aurita, a gelatinous marine invertebrate, by comparing different methodologies and evaluating the influence of tissue type and Proteinase K treatment duration on DNA yield. Method: A total of 48 different combinations were tested using three extraction methods (salting-out, kit-based, and Trizol-based) across four tissue types (gonads, tentacles, epidermis, and gastrodermis). Each method was applied with varying Proteinase K incubation times (0, 1, 3, and 24 hours) to assess their impact on DNA yield and integrity. Findings: Gonadal tissues consistently provided the highest DNA concentrations, likely due to their relatively high cellular density and structural robustness. Among the methods, the salting-out protocol yielded superior results, particularly with 1–3 hours of Proteinase K incubation. In contrast, highly hydrated tissues such as epidermis and gastrodermis led to underestimated yields due to distorted wet weight measurements. Conclusion: Tissue selection, enzymatic treatment time, and extraction methodology critically affect DNA recovery from jellyfish. The salting-out method, when applied to gonadal tissue with moderate Proteinase K incubation, is recommended for efficient and reliable DNA isolation from Aurelia aurita.

Keywords

• DNA isolation
• DNA extraction
• Aurelia aurita
• salting-out method
• DNA yield

Supporting Institution

Scientific Research Projects Coordination Unit (BAP) of Istanbul University

Project Number

FLO-2024-40848

Ethical Statement

Not applicable.

Thanks

Dr. Habeş Bilal AYDEMİR and Dr. Esra SU

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