Title: Development, optimisation and single laboratory validation of an event-specific real-time PCR method for the detection and quantification of Golden Rice 2 using a novel taxon-specific assay
Authors: JACCHIA SARANARDINI ELENASAVINI CristianPETRILLO MAUROANGERS ALEXANDRESHIM Jung-HyunTRIJATMIKO KurniawanKREYSA JOACHIMMAZZARA Marco
Citation: JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY vol. 63 no. 6 p. 1711-1721
Publisher: AMER CHEMICAL SOC
Publication Year: 2015
JRC N°: JRC91800
ISSN: 0021-8561
URI: http://pubs.acs.org/doi/abs/10.1021/jf505516y
http://publications.jrc.ec.europa.eu/repository/handle/JRC91800
DOI: 10.1021/jf505516y
Type: Articles in periodicals and books
Abstract: In this study, we developed, optimized, and in-house validated a real-time PCR method for the event-specific detection and quantification of Golden Rice 2, a genetically modified rice with provitamin A in the grain. We optimized and evaluated the performance of the taxon (targeting rice Phospholipase D α2 gene)- and event (targeting the 3′ insert-to-plant DNA junction)-specific assays that compose the method as independent modules, using haploid genome equivalents as unit of measurement. We verified the specificity of the two real-time PCR assays and determined their dynamic range, limit of quantification, limit of detection, and robustness. We also confirmed that the taxon-specific DNA sequence is present in single copy in the rice genome and verified its stability of amplification across 132 rice varieties. A relative quantification experiment evidenced the correct performance of the two assays when used in combination.
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