İşlenmiş gıdalar için GDO tespitinde modifiye DNA izolasyon yöntemlerinin karşılaştırılması

Öz

İşlenmiş gıda numunelerindeki yüksek oranda bozulmuş DNA içeriği, GDO'ların tespitinde verimi sınırlamaktadır. Genel olarak, literatürde transgenik bitki tohumlarından veya ham maddelerden geleneksel DNA izolasyon teknikleri mevcuttur, ancak işlenmiş gıda örneklerinden DNA izolasyon teknikleri ile ilgili çalışmalar sınırlıdır. Ayrıca, birçok işlenmiş gıda ürünü, çok sayıda karmaşık bitki veya hayvan kaynaklarından elde edilen genomik DNA içerir. Bu çalışma bisküvi, kek, kraker, mısır cipsi ve un gibi işlenmiş gıdaların yüksek kaliteli DNA izolasyonu için bazı faydalı modifikasyonlar önermektedir. Bu amaçla, işlenmiş gıdaları incelemek için GDO analizinin ilk adımı olan gıda örneklerinden yüksek moleküler ağırlıklı ve kaliteli DNA elde etmek için izolasyon protokolleri araştırılmıştır. Elde edilen DNA örneklerinden beklenen uzunluktaki hedef organizmanın gen bölgesini kontrol etmek ve bunları DNA izolasyon verimleriyle karşılaştırmak için soya fasulyesi ve mısıra özgü primerler ile PCR tespiti yapılmıştır. Test edilen dört izolasyon yöntemi arasında, değiştirilmiş Wizard-CTAB yöntemi, test edilen gıda ürünlerinin çoğunda daha iyi sonuçlar vermiştir. Sonuçlar, modifiye Wizard-CTAB'nin mısır ve soya fasulyesine özgü genler ve GDO tespiti ile ilgili çalışmalar için farklı gıda ürünlerinde seçim için kullanılabileceğini göstermiştir.

Anahtar Kelimeler

• DNA izolasyonu,
• genetiği değiştirilmiş organizma (GDO)
• lektin
• nükleik asit eksraksiyonu
• PCR tespiti

Proje Numarası

03-D-21

Comparison of Modified DNA Isolation Methods for the Detection of GMO in Processed Foods

Öz

The highly degraded DNA content in processed food samples results in limited efficiency in detecting GMOs. Generally, conventional DNA isolation techniques from transgenic plant seeds or raw materials are available in the literature, but studies on DNA isolation techniques from processed food samples are more limited. Also, many processed food products contain genomic DNA from numerous complex plants or animal sources. This study proposed some beneficial modifications for high-quality DNA isolation of processed foods such as biscuits, cakes, crackers, corn chips, and flours. For this purpose, isolation protocols were investigated to obtain high molecular weight and quality DNA from food samples, the first step in GMO analysis to study processed foods. PCR detection was performed with soybean and corn-specific primers to control the gene region of the target organism of expected length from the obtained DNA samples and compare them with DNA isolation yields. Among the four isolation methods tested, the modified Wizard-CTAB method showed better results in most of the tested food products. Results showed that the modified Wizard-CTAB could be used for selection can be made in different food products for studies on corn and soybean specific genes and GMO detection.

Anahtar Kelimeler

• DNA isolation
• genetically modified food (GMO)
• lectin
• nucleic acid extraction
• PCR detection

Destekleyen Kurum

Karamanoglu Mehmetbey University Scientific Research Projects Commission

Proje Numarası

03-D-21

Teşekkür

This study was supported by Karamanoglu Mehmetbey University Scientific Research Projects Commission (Project No: 03-D-21). In this article, data belonging to the study of Begum TERZI AKSOY's doctoral thesis were used.

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