Endogenous Reference Genes and Their Quantitative Real-Time PCR Assays for Genetically Modified Bread Wheat (Triticum aestivum L.) Detection

2017 ◽  
pp. 259-268
Author(s):  
Litao Yang ◽  
Sheng Quan ◽  
Dabing Zhang
Keyword(s):  
Triticum Aestivum ◽  
Real Time ◽  
Bread Wheat ◽  
Real Time Pcr ◽  
Reference Genes ◽  
Genetically Modified ◽  
Triticum Aestivum L ◽  
Endogenous Reference ◽  
Pcr Assays ◽  
Endogenous Reference Genes

scholarly journals Evaluation of Four Endogenous Reference Genes and Their Real-Time PCR Assays for Common Wheat Quantification in GMOs Detection

PLoS ONE ◽  
2013 ◽  
Vol 8 (9) ◽  
pp. e75850 ◽  
Author(s):  
Huali Huang ◽  
Fang Cheng ◽  
Ruoan Wang ◽  
Dabing Zhang ◽  
Litao Yang
Keyword(s):  
Real Time ◽  
Common Wheat ◽  
Real Time Pcr ◽  
Reference Genes ◽  
Endogenous Reference ◽  
Pcr Assays ◽  
Endogenous Reference Genes

scholarly journals SYBR®Green qPCR methods for detection of endogenous reference genes in commodity crops: a step ahead in combinatory screening of genetically modified crops in food and feed products

2011 ◽  
Vol 232 (3) ◽  
pp. 485-496 ◽  
Author(s):  
E. Guillaume Mbongolo Mbella ◽  
Antoon Lievens ◽  
Elodie Barbau-Piednoir ◽  
Myriam Sneyers ◽  
Amaya Leunda-Casi ◽  
...  
Keyword(s):  
Reference Genes ◽  
Genetically Modified ◽  
Genetically Modified Crops ◽  
Food And Feed ◽  
Endogenous Reference ◽  
Endogenous Reference Genes

scholarly journals Real-Time Polymerase Chain Reaction-Based Approach for Quantification of the pat Gene in the T25 Zea mays Event

2004 ◽  
Vol 87 (6) ◽  
pp. 1342-1355 ◽  
Author(s):  
Florian Weighardt ◽  
Cristina Barbati ◽  
Claudia Paoletti ◽  
Maddalena Querci ◽  
Simon Kay ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Zea Mays ◽  
Real Time ◽  
Real Time Pcr ◽  
Reference Genes ◽  
Genetically Modified ◽  
Mean Value ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Gmo Quantification

Abstract In Europe, a growing interest for reliable techniques for the quantification of genetically modified component(s) of food matrixes is arising from the need to comply with the European legislative framework on novel food products. Real-time polymerase chain reaction (PCR) is currently the most powerful technique for the quantification of specific nucleic acid sequences. Several real-time PCR methodologies based on different molecular principles have been developed for this purpose. The most frequently used approach in the field of genetically modified organism (GMO) quantification in food or feed samples is based on the 5′-3′-exonuclease activity of Taq DNA polymerase on specific degradation probes (TaqMan® principle). A novel approach was developed for the establishment of a TaqMan quantification system assessing GMO contents around the 1% threshold stipulated under European Union (EU) legislation for the labeling of food products. The Zea mays T25eliteeventwas chosen as a model for the development of the novel GMO quantification approach. The most innovative aspect of the system is represented by the use of sequences cloned in plasmids as reference standards. In the field of GMO quantification, plasmids are an easy to use, cheap, and reliable alternative to Certified Reference Materials (CRMs), which are only available for a few of the GMOs authorized in Europe, have a relatively high production cost, and require further processing to be suitable for analysis. Strengths and weaknesses of the use of novel plasmid-based standards are addressed in detail. In addition, the quantification system was designed to avoid the use of a reference gene (e.g., a single copy, species-specific gene) as normalizer, i.e., to perform a GMO quantification based on an absolute instead of a relative measurement. In fact, experimental evidences show that the use of reference genes adds variability to the measurement system because a second independent real-time PCR-based measurement must be performed. Moreover, for some reference genes no sufficient information on copy number in and among genomes of different lines is available, making adequate quantification difficult. Once developed, the method was subsequently validated according to IUPAC and ISO 5725 guidelines. Thirteen laboratories from 8 EU countries participated in the trial. Eleven laboratories provided results complying with the predefined study requirements. Repeatability (RSDr)values ranged from 8.7 to 15.9%, with a mean value of 12%. Reproducibility (RSDR) values ranged from 16.3 to 25.5%, with a mean value of 21%. Following Codex Alimentarius Committee guidelines, both the limits of detection and quantitation were determined to be <0.1%.

Collaborative trial validation of cry1Ab/Ac and Pubi-cry TaqMan-based real-time PCR assays for detection of DNA derived from genetically modified Bt plant products

2015 ◽  
Vol 20 (2) ◽  
pp. 85-96 ◽  
Author(s):  
Lutz Grohmann ◽  
Ralf Reiting ◽  
Dietrich Mäde ◽  
Steffen Uhlig ◽  
Kirsten Simon ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Genetically Modified ◽  
Plant Products ◽  
Collaborative Trial ◽  
Pcr Assays
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