Abstract
Background Brazil is the second largest producer of genetically modified (GM) crops with 37 GM crop varieties approved by CTNBio for commercialization. One of these varieties is the Embrapa 5.1 common bean, resistant to the bean golden mosaic virus (BGMV). The Embrapa 5.1 event was developed using the RNAi concept to induce the silencing of the AC1 viral gene [1]. Labelling of food products containing GMOs in Brazil is mandatory [2], so methods for quantification of GMOs are required. Real time PCR methods have been used as the gold standard for GMO quantification. In a previous work, we have developed a construct-specific PCR detection assay for Embrapa 5.1 [3]. In this study we developed an event-specific PCR assay for Embrapa 5.1.
Highlights
Brazil is the second largest producer of genetically modified (GM) crops with 37 GM crop varieties approved by CTNBio for commercialization
Genomic DNA was extracted from leaves using two different protocols, a CTAB method and the DNeasy Plant Mini Kit with modifications [3]
The endogenous reference presented an efficiency of 96% and a limit of detection (LOD) of 10 genome copies
Summary
Brazil is the second largest producer of genetically modified (GM) crops with 37 GM crop varieties approved by CTNBio for commercialization. Methods Genomic DNA was extracted from leaves using two different protocols, a CTAB method and the DNeasy Plant Mini Kit with modifications [3]. Quantification of the endogenous reference was performed using primers and Taqman probe targeting the lectin gene present in common bean (Phaseolus vulgaris) genome. The reaction parameters efficiency and limit of detection (LOD) were determined using conventional and GM varieties Perola and Pontal by 10-fold serial dilution of the genomic DNA ranging from 105 to 100 genome copies.
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