Using plasmid reference materials for genetically modified organisms analysis and their verification with inter-laboratory comparison test

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Tezin Türü: Yüksek Lisans

Tezin Yürütüldüğü Kurum: Orta Doğu Teknik Üniversitesi, Fen Bilimleri Enstitüsü, Fen Bilimleri Enstitüsü, Türkiye

Tezin Onay Tarihi: 2016

Öğrenci: TUĞÇE CEREN TUĞRUL

Danışman: AYŞE MERAL YÜCEL

Özet:

Genetically modified organisms (GMOs) have been strictly controlled with legislative regulation in many countries. In this study, plasmid reference materials (PRMs) for identification and quantification of GMOs had been developed and the verification of PRMs was done to be done with interlaboratory comparison test (ICT). PRMs made of plasmid DNA that were used for developing GMO analysis method. In this study, four PRMs were developed and their optimization was achieved with quantitative polymerase chain reaction (Q-PCR) and SYBR green I method. Commercially valuable maize and soybean are frequently subject to genetic modification (GM). ADH1 for maize and Lectin for soybean were chosen as an endogenous gene. Moreover, BT11 event in maize and RR event in soybean were chosen as genetic modification. Overall, there are two PRMs for each plant to determine amount of plant and quantify GM by GMO analysis. vi The study has 3 main steps. In the first step, development and optimization of special method for PRMs was achieved. PRMs were subjected to single laboratory test and measurement uncertainty as second step. Considering results, third step was done with ICT. PRM validation test sets were prepared and send to ten different public and private food analysis laboratories. PRMs were analyzed and results were calculated statistically. PRMs are equal to certified reference materials that are imported with high price from Europe. This is the first study of ICT with reference material in Turkey. In addition, methods union and standardization of result evaluation about GMO analysis was identified