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
Öktem, Hüseyin Avni
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 probe and a fluorescein-tagged signal probe, which are partially complementary to the p35S oligonucleotide. We used poly-l-lysine-coated glass slides as support material, on which capture probes were immobilized by a heterobifunctional cross-linker. The comparative results of optimization studies including cross-linker types probe concentrations and hybridization conditions (sequence, temperature and duration) were reported. An optimum hybridization signal was obtained with a 32.5 C-0 cross-linker, 10 mu M capture and 20 mu M signal probe concentrations, respectively. A relatively short hybridization time (2.5 h) provided reproducible array signals. No significant effect of hybridization sequence on the fluorescence intensity was observed. The described platform can specifically detect label-free transgenic sequences with a target of 0.01 mu M concentration, while the optimized system exhibits great potential for the application of different GMO target sequences (p35S, tNOS, bar and cry) to multiplex array formats.


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 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...
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.
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 ...
Citation Formats
S. Cansiz et al., “A sandwich-type DNA array platform for detection of GM targets in multiplex assay,” EUROPEAN FOOD RESEARCH AND TECHNOLOGY, pp. 429–437, 2012, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/30828.