GDO analizlerinde kullanılan RM izolatlarının kontrollü muhafaza koşullarında tekrar kullanım olanaklarının araştırılması
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Dosyalar
Tarih
2020
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Bursa Teknik Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Genetiği değiştirilmiş organizmalar (GDO) avantaj ve dezavantajları tartışılmakla beraber potansiyel yararları nedeniyle halen yaygın olarak üretilen ürünlerdir. Ürünlerde GDO varlığının tespiti kalitatif tarama ve tiplendirme analizleri ile belirlenirken, miktar analizleri kantitatif testlerle yapılmaktadır. GDO analizlerinden tiplendirme analizlerinde pozitif kontrol olarak ve miktar analizlerinde kalibrasyon eğrisi hazırlamada sertifikalı referans maddeler (CRM) kullanılmaktadır. Analizlerde kullanılan CRM'ler her seferinde taze olarak hazırlanmaktadır. Ancak bu materyallerin çok pahalı olması ve hazırlanan miktarın tek seferde kullanılamaması bu analizi yapan laboratuvarlar için önemli bir sorun oluşturmaktadır. Bu soruna çözüm olarak analizlerden geriye kalan CRM izolatlarının tekrar kullanılabilme olanaklarının belirlenmesi önem arz etmektedir. Deoksiribo nükleik asit (DNA)'e uygulanan dondur-çözdür işlemlerinde DNA'nın degrade olup olmadığı ile ilgili bitkisel ürünlerde yeterli bilimsel veri bulunmadığı fark edilmiş olup çalışmaların ise daha çok parazit, virüs ve kan örnekleri ile sınırlı olduğu görülmüştür. Planlanan bu çalışma ile bitkisel ürünlerde GDO miktar analizlerinde kullanılan CRM'lerden elde edilen DNA izolatlarında kontrollü muhafaza koşullarında dondur-çözdür uygulaması ile meydana gelebilecek degradasyonun belirlenmesi amaçlanmıştır. Bu amaçla soya (DP-305423-1) ve mısır (DAS-40278-9) genini içeren CRM'lerden Eurofins Genespin Kit Metodu ile DNA izolasyonu yapılmış, elde edilen izolatlara belirli sıklıklarla dondurma (-20±2°C) ve laboratuvar koşullarında (22±2°C) çözdürme işlemleri uygulanmıştır. Avrupa Birliği Referans Laboratuvarı (EURL) metodları kullanılarak Applied Biosystems 7500 Fast Real time PCR System cihazında GDO miktar analizi yapılmış, elde edilen veriler SPSS paket programına göre istatistiki olarak değerlendirilerek dondur-çözdür uygulamasının çalışılan CRM izolatları üzerinde etkisi önemsiz bulunmuştur.
Although the advantages and disadvantages of genetically modified organisms (GMOs) are discussed, they are still widely produced due to their potential benefits. Detection of the presence of GMO in the products is determined by qualitative screening and typing analyzes, while quantity analyzes are carried out by quantitative tests. Certified reference materials (CRM) are used as positive controls in typing analyzes from GMO analyzes and in calibration curves in quantity analyzes. The CRMs used in the analyzes are prepared fresh each time. However, the fact that these materials are very expensive and that the prepared amount cannot be used at one time poses an important problem for the laboratories performing this analysis. As a solution to this problem, it is important to determine the possibilities of reuse of the CRM isolates remaining from the analyzes. In the freeze-thaw processes applied to deoxyribo nucleic acid (DNA), it was found that there was not enough scientific data on plant products about the degradation of DNA and the studies were found to be mostly limited to parasites, viruses and blood samples. The aim of this study was to determine the degradation of freeze-thaw application of DNA isolates obtained from CRMs used in GMO quantity analysis of plant products under freeze-thaw application under controlled storage conditions. For this purpose, DNA isolation was performed by using Eurofins Genespin Kit Method from CRMs containing soy (DP-305423-1) and corn (DAS-40278-9) gene, and freeze (-20 ± 2 ° C) thawing (22 ± 2 ° C) processes were applied to the obtained isolates at certain frequencies. GMO Quantitative analysis was performed on Applied Biosystems 7500 Fast Real Time PCR System using the European Union Reference Laboratory (EURL) methods. The obtained data were evaluated statistically according to SPSS package program and the effect of freeze-thaw application on the CRM isolates studied was insignificant.
Although the advantages and disadvantages of genetically modified organisms (GMOs) are discussed, they are still widely produced due to their potential benefits. Detection of the presence of GMO in the products is determined by qualitative screening and typing analyzes, while quantity analyzes are carried out by quantitative tests. Certified reference materials (CRM) are used as positive controls in typing analyzes from GMO analyzes and in calibration curves in quantity analyzes. The CRMs used in the analyzes are prepared fresh each time. However, the fact that these materials are very expensive and that the prepared amount cannot be used at one time poses an important problem for the laboratories performing this analysis. As a solution to this problem, it is important to determine the possibilities of reuse of the CRM isolates remaining from the analyzes. In the freeze-thaw processes applied to deoxyribo nucleic acid (DNA), it was found that there was not enough scientific data on plant products about the degradation of DNA and the studies were found to be mostly limited to parasites, viruses and blood samples. The aim of this study was to determine the degradation of freeze-thaw application of DNA isolates obtained from CRMs used in GMO quantity analysis of plant products under freeze-thaw application under controlled storage conditions. For this purpose, DNA isolation was performed by using Eurofins Genespin Kit Method from CRMs containing soy (DP-305423-1) and corn (DAS-40278-9) gene, and freeze (-20 ± 2 ° C) thawing (22 ± 2 ° C) processes were applied to the obtained isolates at certain frequencies. GMO Quantitative analysis was performed on Applied Biosystems 7500 Fast Real Time PCR System using the European Union Reference Laboratory (EURL) methods. The obtained data were evaluated statistically according to SPSS package program and the effect of freeze-thaw application on the CRM isolates studied was insignificant.
Açıklama
Anahtar Kelimeler
Biyoteknoloji, Biotechnology
