Simultaneous Detection of Recombinant DNA Segments Introduced into Genetically Modified Crops with Multiplex Ligase Chain Reaction Coupled with Multiplex Polymerase Chain Reaction

2009 ◽  
Vol 57 (7) ◽  
pp. 2640-2646 ◽  
Author(s):  
Junichi Mano ◽  
Taichi Oguchi ◽  
Hiroshi Akiyama ◽  
Reiko Teshima ◽  
Akihiro Hino ◽  
...  
2009 ◽  
Vol 92 (3) ◽  
pp. 765-772 ◽  
Author(s):  
Anna Nadal ◽  
Teresa Esteve ◽  
Maria Pla

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.


2016 ◽  
Vol 32 (6) ◽  
pp. 575-579 ◽  
Author(s):  
In Jeong Kang ◽  
Mi-Hyung Kang ◽  
Tae-Hwan Noh ◽  
Hyeong Kwon Shim ◽  
Dong Bum Shin ◽  
...  

Crop Science ◽  
2010 ◽  
Vol 50 (5) ◽  
pp. 1961-1966 ◽  
Author(s):  
Baigalmaa Jigden ◽  
Hongtao Wang ◽  
Yeon-Ju Kim ◽  
Jong-Hun Noh ◽  
Choijamts Gotov ◽  
...  

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