A new dual plasmid calibrator for the quantification of the construct specific GM canola Oxy-235 with duplex real-time PCR
| Autor: | Khaled Said, Maher Chaouachi, Nesrine Nabi, Mohamed Salem Zellama, Ahmed Ben Hafsa |
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| Rok vydání: | 2013 |
| Předmět: |
food.ingredient
DNA Plant Analytical chemistry Real-Time Polymerase Chain Reaction Analytical Chemistry food media_common.cataloged_instance European union Canola media_common Mathematics Detection limit Reproducibility Chromatography Brassica napus Reproducibility of Results General Medicine Repeatability Reference Standards Plants Genetically Modified Real-time polymerase chain reaction Certified reference materials Duplex (building) Calibration Food Science Plasmids |
| Zdroj: | Food chemistry. 145 |
| ISSN: | 1873-7072 |
| Popis: | To overcome the difficulties of obtaining the Certified Reference Material (CRM) and according to the key documents of the European Union Reference Laboratory (EU-RL), a new standard reference molecule containing the construct specific of the canola event Oxy-235 (3′-junction Nitrilase/Tnos) and the canola endogenous reference gene (acety-CoA-carboxylase) was constructed and used for duplex real-time quantitative analysis. The limits of detection (LOD) were less than 5 Haploid Genome Copy (HGC) and the limits of quantification (LOQ) were about 10 HGC. Furthermore, mixed GM and non-GM canola samples were analysed with duplex QRT-PCR to evaluate the performance criteria as required for validation procedures in the EU-RL, namely, the precision and the accuracy. The accuracy expressed as bias ranged from 2% to 10% and the precision (repeatability and reproducibility) expressed as the RSD r and RSD R was from 2.2 to 5.12 and 2.15 to 5.46 respectively. All these indicated that the developed construct specific method and the reference molecule are suitable for the identification and the quantification of the canola event Oxy-235. |
| Databáze: | OpenAIRE |
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