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

Tuğrul, Tuğçe Ceren
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


Development of reference materials for GMO detection /
Keskin, Batuhan Birol; Öktem, Hüseyin Avni; Department of Biotechnology (2014)
The reliable detection and quantification of Genetically Modified Organisms (GMO) is strongly dependent on validated methods as well as calibration systems. Today, Certified Reference Materials are used as reliable source of template DNA in quantitative real-time PCR assays constructing validated methods for routine analysis of GMOs. In addition to that, obtaining and assessing plasmids for use as real-time PCR standards and positive PCR controls are increasingly used methodologies. To enforce the labeling ...
Development of a loop mediated isothermal amplification (LAMP) based detection platform for genetically modified organism (GMO) detection
Moğol, Ayça Nazlı; Öktem, Hüseyin Avni; Department of Biotechnology (2019)
Genetically modified organisms (GMOs) are being widely used worldwide. Every country has a different legislation regarding the allowed events and GMO levels. This creates the great need for GMO detection. In this study, LAMP assay was used for GMO detection owing to its high sensitivity with the genetically modified organisms; Bt11 maize, GT73 Roundup Ready canola, and transgenic Nicotiana tabacum, targeting the sequences most commonly used in GMO constructions; 35S promoter and Figwort mosaic virus (FMV) s...
A sandwich-type DNA array platform for detection of GM targets in multiplex assay
Cansiz, Sena; Özen, Can; Bayrac, Ceren; Bayrac, A. Tahir; Gül, Fatma; Kavruk, Murat; Yilmaz, Remziye; EYİDOĞAN, FÜSUN; Öktem, Hüseyin Avni (2012-09-01)
Over the last decade, array detection has been developed to qualitatively assess the presence of genetically modified organisms (GMOs). To date, DNA array systems have the highest capabilities as a result of GMOs analysis. We describe the construction of an array platform in the sandwich hybridization format for the detection of transgenic promoter of Cauliflower mosaic virus (CaMV; p35S). Sequence-specific signal development has been achieved by a sandwich complex composed of a surface immobilized capture ...
Development of analysis methods for cry1ac and sam-k gene lines in tomato using pcr and real-time pcr
Uygun, Sahra; Gültekin, Güzin Candan; Department of Biotechnology (2010)
Genetically modified organisms are entering the human diet in all over the world. In order to have transparency in the foods that are being consumed, there is a need to trace the genetically modified organisms (GMOs) in the market and consequently this need brings the necessity of analytical methods that are capable of detecting, identifying and quantifying the transgenic events. These analytical methods also form the basis of the labeling regulations that are tried to be formed regarding GMOs. The main aim...
Qualitative detection of GM maize (Bt 11) in food and feed sold commercially in Turkey by PCR based methods
Gültekin, Güzin Candan; Yilmaz, Remziye (2011-01-01)
To regulate the presence of genetically modified organisms (GMOs) in crops, foods and food ingredients, the development of reliable and sensitive methods for their detection is necessary. In the present study, products derived from GM maize in the Turkish market were monitored using methods to detect the presence of CaMV 35S promoter, nos terminator and Bt 11 maize. The results demonstrated, for the first time, the presence of GM maize in Turkish food and feed products qualitatively.
Citation Formats
T. C. Tuğrul, “Using plasmid reference materials for genetically modified organisms analysis and their verification with inter-laboratory comparison test,” M.S. - Master of Science, Middle East Technical University, 2016.