Development of a loop mediated isothermal amplification (LAMP) based detection platform for genetically modified organism (GMO) detection

Download

index.pdf

Date

2019

Author

Moğol, Ayça Nazlı

Metadata

Show full item record

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Item Usage Stats

320
views

340
downloads

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) sequences. Optimal conditions for the LAMP assay have been determined and with these conditions, sensitivity and specificity tests of the LAMP assays of 35S promoter were done using Bt11 maize and Nicotiana tabacum. The sensitivity and specificity tests of the LAMP assays of FMV were done using GT73 Roundup Ready canola. Hydroxy naphthol blue dye was used for monitoring of the LAMP products in addition to the agarose gel (1.5%) electrophoresis. The sensitivity tests showed that GMO detection was possible for as low as 1 double stranded genomic DNA copy for both 35S promoter and FMV sequences. The specificity tests showed that both of the primer sets (35S promoter and FMV) used in this study were highly specific and their specificity was not affected by the presence of a foreign DNA. Rapid DNA extraction techniques were also tested with the LAMP assay, and it was found that DNA extraction step could be finished as short as 5 minutes. Finally, lyophilized LAMP assay successfully detected 35S promoter sequence in Bt11 maize.

Subject Keywords

Genetics., GMO, LAMP, Sensitivity Test, Specificity Test, Hydroxy Naphthol Blue

URI

http://etd.lib.metu.edu.tr/upload/12623362/index.pdf
https://hdl.handle.net/11511/43762

Collections

Graduate School of Natural and Applied Sciences, Thesis

Suggestions

OpenMETU
Core

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...
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...
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...
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

A. N. Moğol, “Development of a loop mediated isothermal amplification (LAMP) based detection platform for genetically modified organism (GMO) detection,” Thesis (M.S.) -- Graduate School of Natural and Applied Sciences. Biotechnology., Middle East Technical University, 2019.