Detection of exogenous genes in genetically modified plants with multiplex polymerase chain reaction

2001 ◽  
Vol 19 (4) ◽  
pp. 289-298 ◽  
Author(s):  
Zhen Tao ◽  
Xing-Feng Cai ◽  
Sheng-Li Yang ◽  
Yi Gong
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Polymerase Chain Reaction ◽  
Genetically Modified ◽  
Genetically Modified Plants ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Exogenous Genes

scholarly journals Multiplex Polymerase Chain Reaction-Capillary Gel Electrophoresis: A Promising Tool for GMO ScreeningAssay for Simultaneous Detection of Five Genetically Modified Cotton Events and Species

2009 ◽  
Vol 92 (3) ◽  
pp. 765-772 ◽  
Author(s):  
Anna Nadal ◽  
Teresa Esteve ◽  
Maria Pla
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Gel Electrophoresis ◽  
Multiplex Polymerase Chain Reaction ◽  
Genetically Modified ◽  
Simultaneous Detection ◽  
Chain Reaction ◽  
Capillary Gel Electrophoresis ◽  
Cotton Species ◽  
Polymerase Chain

Abstract A multiplex polymerase chain reaction assay coupled to capillary gel electrophoresis for amplicon identification by size and color (multiplex PCR-CGE-SC) was developed for simultaneous detection of cotton species and 5 events of genetically modified (GM) cotton. Validated real-time-PCR reactions targeting Bollgard, Bollgard II, Roundup Ready, 3006-210-23, and 281-24-236 junction sequences, and the cotton reference gene acp1 were adapted to detect more than half of the European Union-approved individual or stacked GM cotton events in one reaction. The assay was fully specific (<1.7 of false classification rate), with limit of detection values of 0.1 for each event, which were also achieved with simulated mixtures at different relative percentages of targets. The assay was further combined with a second multiplex PCR-CGE-SC assay to allow simultaneous detection of 6 cotton and 5 maize targets (two endogenous genes and 9 GM events) in two multiplex PCRs and a single CGE, making the approach more economic. Besides allowing simultaneous detection of many targets with adequate specificity and sensitivity, the multiplex PCR-CGE-SC approach has high throughput and automation capabilities, while keeping a very simple protocol, e.g., amplification and labeling in one step. Thus, it is an easy and inexpensive tool for initial screening, to be complemented with quantitative assays if necessary.

scholarly journals Detection of genetically modified plants using LAMP (loop-mediated amplification) technologies

2021 ◽  
Vol 17 (1) ◽  
pp. 51-59
Author(s):  
B. V. Sorochynskyi
Keyword(s):  
Polymerase Chain Reaction ◽  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Genetically Modified Plants ◽  
Chain Reaction ◽  
Plant Materials ◽  
Advantages And Disadvantages ◽  
Current State ◽  
Polymerase Chain

Purpose. Analysis of the current state and experience on the loop-mediated amplification (LAMP) use to detect genetically modified plants. Methods. Literature search and analysis. Results. General information on the current state and use of the genetically modified plants is provided. Despite the wide distribution of genetically modified plants, the attitude towards them in society continues to remain somewhat wary. About 50 countries have introduced mandatory labeling of GM feed and products, provided that their content exceeds a certain threshold. In order to meet labeling requirements, effective and sensitive methods for detecting known genetic modifications in a variety of plant materials, food products and animal feed must be developed and standardized. The most common approaches to the detection of genetically modified organisms (GMOs) are the determination of specific proteins synthesized in transgenic plants and the detection of new introduced genes. Methods for the determination of GMOs based on the analysis of nucleic acids are more common, since such methods have greater sensitivity and specificity than the analysis of protein composition. Polymerase chain reaction (PCR) method is the main method of nucleic acid analysis, which is now wide used for the detection of GMOs. Loop-mediated amplification (LAMP), which can occur at a constant temperature and therefore does not require the use of expensive equipment may be an alternative to the PCR. Scientific articles about the use of the loop-mediated amplification (LAMP) for the detection of genetically modified plants were analyzed. Advantages and disadvantages of the polymerase chain reaction and loop-mediated amplification are compared. Conclusions. The main criteria for applying a method of GMO detection analysis are as follow: its sensitivity, time of reaction, availability and ease to use, cost of reagents and equipment, and the possibility for simultaneous detection of many samples.

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