Elaboration of a Reliable Strategy Based on Real-Time PCR To Characterize Genetically Modified Plantlets and To Evaluate the Efficiency of a Marker Gene Removal in Grape (Vitisspp.)

2009 ◽  
Vol 57 (7) ◽  
pp. 2668-2677 ◽  
Author(s):  
Lorenza Dalla Costa ◽  
Ilaria Vaccari ◽  
Marco Mandolini ◽  
Lucia Martinelli
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Genetically Modified ◽  
Marker Gene

scholarly journals Detection and copy number estimation of the transgenic nucleotide sequences in an unknown GM event of Oryza sativa

2016 ◽  
Vol 69 (4) ◽  
Author(s):  
Ali M. Sajjad ◽  
Tanzeela Bashir ◽  
Shaina Saeed ◽  
Emmad Ahmad
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Copy Number ◽  
Genetically Modified ◽  
Marker Gene ◽  
Experimental Conditions ◽  
Rice Varieties ◽  
Genetically Modified Rice ◽  
Qualitative Detection

The present study was designed to establish a qualitative detection method based on conventional and real time PCR assay to screen the commonly grown rice varieties for the presence of the <em>cry1Ac</em> gene. The detection of genetically modified rice in the screening process would necessitate accurate assay development and precise qualitative PCR tests complying with established procedures for the detection and characterization of transgenes in food grains. Such assay would not only enable the monitoring of transgene flow in local agricultural environment but also the characterization of different plant species produced with this transgene and its regulatory components. Thus, a reliable and quick screening assay was established for the qualitative detection of the transgene along with the promoter and selectable marker gene in genetically modified rice. By conventional PCR, a fragment of 215 bp was amplified with gene specific primers of <em>cry1Ac</em>. Primers for other transgenes such as <em>gna</em> and <em>bar</em> were also employed; however, no amplification was detected. The presence of the <em>p35s</em>, <em>sps</em>, and <em>nptII</em> genes was confirmed by qualitative real-time PCR. The specificity of the respective PCR products was checked through melt peak curve analysis. Sharp and precise melting temperatures indicated the presence of a single kind of PCR product in correspondence to each of the primers used. Moreover, the copy number of <em>cry1Ac</em> was estimated by ∆∆<em>C</em><span><sub>T</sub></span> method. It is proposed that the primer sets and experimental conditions used in this study will be sufficient to meet the requirements for molecular detection and characterization of the <em>cry1Ac</em> transgene and affiliated sequences in sorting out conventional rice varieties from the ones which are genetically modified. It will also help to monitor the ecological flow of these transgenes and other biosafety factors.

scholarly journals Effect of Feeding Cows Genetically Modified Maize on the Bacterial Community in the Bovine Rumen

2007 ◽  
Vol 73 (24) ◽  
pp. 8012-8017 ◽  
Author(s):  
S. Wiedemann ◽  
P. Gürtler ◽  
C. Albrecht
Keyword(s):  
Bacterial Community ◽  
Real Time ◽  
Real Time Pcr ◽  
Genetically Modified ◽  
Genetically Modified Maize ◽  
Bacterial Strains ◽  
Maize Silage ◽  
Bovine Rumen ◽  
Gm Maize ◽  
Effect Of Feeding

ABSTRACT Rumen-cannulated cows (n = 4) were fed successively silage made from either conventional or genetically modified (GM) maize. Results revealed no effects of GM maize on the dynamics of six ruminal bacterial strains (investigated by real-time PCR) compared to the conventional maize silage.

scholarly journals Quantification of Genetically Modified Soybeans Using a Combination of a Capillary-Type Real-Time PCR System and a Plasmid Reference Standard

2006 ◽  
Vol 70 (4) ◽  
pp. 821-827 ◽  
Author(s):  
Akie TOYOTA ◽  
Hiroshi AKIYAMA ◽  
Mitsunori SUGIMURA ◽  
Takahiro WATANABE ◽  
Hiroyuki KIKUCHI ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Genetically Modified ◽  
Reference Standard ◽  
Capillary Type ◽  
Genetically Modified Soybeans

Detection by real-time PCR and pyrosequencing of the cry1Ab and cry1Ac genes introduced in genetically modified (GM) constructs

2017 ◽  
Vol 34 (8) ◽  
pp. 1398-1409 ◽  
Author(s):  
Frederic Debode ◽  
Eric Janssen ◽  
Claude Bragard ◽  
Gilbert Berben
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Genetically Modified

scholarly journals Tendency for Interlaboratory Precision in the GMO Analysis Method Based on Real-Time PCR

2010 ◽  
Vol 93 (2) ◽  
pp. 734-749 ◽  
Author(s):  
Takashi Kodama ◽  
Yasunori Kurosawa ◽  
Kazumi Kitta ◽  
Shigehiro Naito
Keyword(s):  
Standard Deviation ◽  
Real Time ◽  
Dna Extraction ◽  
Real Time Pcr ◽  
Quantitative Methods ◽  
Genetically Modified ◽  
Performance Criterion ◽  
Gm Crops ◽  
Evaluation Procedure ◽  
Method Performance

Abstract The Horwitz curve estimates interlaboratory precision as a function only of concentration, and is frequently used as a method performance criterion in food analysis with chemical methods. The quantitative biochemical methods based on real-time PCR require an analogous criterion to progressively promote method validation. We analyzed the tendency of precision using a simplex real-time PCR technique in 53 collaborative studies of seven genetically modified (GM) crops. Reproducibility standard deviation (SR) and repeatability standard deviation (Sr) of the genetically modified organism (GMO) amount () was more or less independent of GM crops (i.e., maize, soybean, cotton, oilseed rape, potato, sugar beet, and rice) and evaluation procedure steps. Some studies evaluated whole steps consisting of DNA extraction and PCR quantitation, whereas others focused only on the PCR quantitation step by using DNA extraction solutions. Therefore, SR and Sr for GMO amount () are functions only of concentration similar to the Horwitz curve. We proposed SR 0.1971C0.8685 and Sr 0.1478C0.8424, where C is the GMO amount (). We also proposed a method performance index in GMO quantitative methods that is analogous to the Horwitz Ratio.

Two FAST multiplex real-time PCR reactions to assess the presence of genetically modified organisms in food

2019 ◽  
Vol 274 ◽  
pp. 760-765 ◽  
Author(s):  
Geoffrey Cottenet ◽  
Carine Blancpain ◽  
Véronique Sonnard ◽  
Poh Fong Chuah
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Genetically Modified Organisms ◽  
Genetically Modified
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