Development of reference materials for GMO detection /

Keskin, Batuhan Birol
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 regulations of GMOs, the application of DNA plasmids as calibrants is becoming essential for the practical quantification of GMOs. This study reports the construction of plasmids for qualitative screening assay for 35S promoter and NOS terminator as GMO elements, and relative quantification assays in Maize and Soya events, Bt11 and GTS 40-3-2 respectively. Reference GM plasmids provided convenient and reliable positive controls for GM PCR tests. Soya GM event GTS 40-3-2 and endogenous control plasmids assessed within-laboratory assays were resulted to be acceptable in terms of reproducibility standard deviation and repeatability relative standard deviation. As a result of verification and measurement uncertainty data based on single laboratory data, constructed plasmids provided an excellent and economic alternative to plant DNA extractions for positive control material. However, further study is needed showing enhanced amplification efficiency and inter-laboratory verification data for using constructed plasmids as template DNA in validated methods.


Using plasmid reference materials for genetically modified organisms analysis and their verification with inter-laboratory comparison test
Tuğrul, Tuğçe Ceren; Yücel, Ayşe Meral; Yılmaz, Remziye; Department of Biotechnology (2016)
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 re...
Discovering functional interaction patterns in protein-protein interaction networks
Turanalp, Mehmet E.; Can, Tolga (Springer Science and Business Media LLC, 2008-06-11)
Background: In recent years, a considerable amount of research effort has been directed to the analysis of biological networks with the availability of genome-scale networks of genes and/or proteins of an increasing number of organisms. A protein-protein interaction (PPI) network is a particular biological network which represents physical interactions between pairs of proteins of an organism. Major research on PPI networks has focused on understanding the topological organization of PPI networks, evolution...
Development of qcm based dna biosensors for detection of genetically modified organisms
Karamollaoğlu, İrem; Öktem, Hüseyin Avni; Department of Biology (2007)
A great effort has been recently devoted to the development of new devices for the detection of specific sequences of DNA, due to increasing need of label - free, fast, cheap, and miniaturized analytical systems able to detect target sequences for screening purposes, especially in food industry for genetically modified organisms (GMOs). In this study, development of a QCM - based DNA biosensor for the detection of the hybridisation of CaMV 35S promoter sequence (P35S) was investigated. Attention was focused...
Genetic differentiation of Abies equi-trojani (Asch. & Sint. ex Boiss) Mattf. populations from Kazdaǧι, Turkey and the genetic relationship between Turkish firs belonging to the Abies nordmanniana Spach complex
Kaya, Zeki; Neale, David Brian (2008-02-05)
The present study aimed to test the utility of RAPD (randomly amplified polymorphic DNA) and cpSSR (simple sequence repeats) markers for in situ gene conservation programs for fir species, as well as for determining the genetic similarities between the Abies nordmanniana Spach species complex (A. nordmanniana, A. bornmuelleriana Matff., A. equi-trojani (Asch. & Sint. ex Boiss) Mattf.) and between populations of A. equi-trojani, which is a narrow-endemic to Turkey. For this purpose, DNA was extracted and poo...
A rapid method for detection of genetically modified organisms based on magnetic separation and surface-enhanced Raman scattering
GÜVEN, BURCU; BOYACI, İSMAİL HAKKI; TAMER, UĞUR; Çalık, Pınar (Royal Society of Chemistry (RSC), 2012-01-01)
In this study, a new method combining magnetic separation (MS) and surface-enhanced Raman scattering (SERS) was developed to detect genetically modified organisms (GMOs). An oligonucleotide probe which is specific for 35 S DNA target was immobilized onto gold coated magnetic nanospheres to form oligonucleotide-coated nanoparticles. A self assembled monolayer was formed on gold nanorods using 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB) and the second probe of the 35 S DNA target was immobilized on the activa...
Citation Formats
B. B. Keskin, “Development of reference materials for GMO detection /,” M.S. - Master of Science, Middle East Technical University, 2014.