Event-specific qualitative and quantitative polymerase chain reaction methods for detection of insect-resistant genetically modified Chinese cabbage based on the 3′-junction of the insertion site

2012 ◽  
Vol 55 (3) ◽  
pp. 367-375 ◽  
Author(s):  
Kong-Sik Shin ◽  
Myung-Ho Lim ◽  
Hee-Jong Woo ◽  
Sun-Hyung Lim ◽  
Hong-Il Ahn ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chinese Cabbage ◽  
Genetically Modified ◽  
Quantitative Polymerase Chain Reaction ◽  
Insertion Site ◽  
Chain Reaction ◽  
Qualitative And Quantitative ◽  
Polymerase Chain ◽  
Insect Resistant

Event-specific quantitative polymerase chain reaction methods for detection of double-herbicide-resistant genetically modified corn MON 87419 based on the 3′-junction of the insertion site

2021 ◽  
Author(s):  
Likun Long ◽  
Wei Yan ◽  
Congcong Li ◽  
Liming Dong ◽  
Na Liu ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Genetically Modified ◽  
Quantitative Polymerase Chain Reaction ◽  
Insertion Site ◽  
Chain Reaction ◽  
Herbicide Resistant ◽  
Qualitative Polymerase Chain Reaction ◽  
Relative Standard ◽  
Junction Site ◽  
Polymerase Chain

ABSTRACT MON 87419 was one of the new transgenic corn events developed in US with the trait of herbicide resistance to both dicamba and glyphosate. To monitor unintended release of genetically modified organism in the future, as well as to meet GM-labeling requirements, it is requisite to develop a reliable method for the detection and quantification of MON 87419, an event-specific primer pair was designed to amplify the 3′-junction site between the endogenous genome sequence and the transferred DNA of GM event MON 87419, amplicons of desired size were produced by qualitative polymerase chain reaction (PCR) assay. For the validation of this quantitative method, the mixed samples containing 10%, 1%, and 0.1% MON 87419 ingredient were quantified. The precisions were expressed as relative standard deviations, deviated by 7.87%, 12.94%, and 19.98%, respectively. These results clearly demonstrate that the PCR methods we developed herein can be used for event-specific quantitative testing of the double-herbicide-resistant corn MON 87419.

scholarly journals Detection of Genetic Modification in Cured Tobacco Leaf: Proficiency Testing Using Polymerase Chain Reaction-Based Methods

2006 ◽  
Vol 89 (3) ◽  
pp. 693-707
Author(s):  
Ferruccio Gadani ◽  
Martin Ward ◽  
Sue Black ◽  
Neil Harris ◽  
David McDowell ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Proficiency Testing ◽  
Quantitative Methods ◽  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Cauliflower Mosaic Virus ◽  
Chain Reaction ◽  
Tobacco Leaf ◽  
Qualitative And Quantitative ◽  
Polymerase Chain

Abstract The Cooperation Centre for Scientific Research Relative to Tobacco (CORESTA; Paris, France) Task Force Genetically Modified TobaccoDetection Methods investigated the performance of qualitative and quantitative methods based on the polymerase chain reaction (PCR) for the detection and quantitation of genetically modified (GM) tobacco. In the 4 successful rounds of proficiency testing, the cauliflower mosaic virus 35S RNA promoter (CaMV 35S) and the Agrobacterium tumefaciens nopaline synthase terminator (NOS) were selected as target sequences. Blind-coded reference materials containing from 0.1 to 5.0% and from 0.15 to 4% GM tobacco were used in 2 rounds of qualitative and quantitative PCR, respectively. Eighteen laboratories from 10 countries participated in this study. Considering all methods and 2 rounds, the different laboratories were able to detect GM tobacco at the 0.1% level in 46 out of 58 tests in qualitative assays. The results of the proficiency test indicate that both end point screening and real-time quantitative methods are suitable for the detection of genetically modified organisms in tobacco leaf samples having a GM content of 0.1% or higher. The CORESTA proficiency study represents a first step towards the interlaboratory evaluation of accuracy and precision of PCR-based GM tobacco detection, which may lead to the harmonization of analytical procedures and to the enhancement of comparability of testing results produced by different laboratories.

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