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
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.
JACCHIA Sara;
NARDINI Elena;
SAVINI Cristian;
PETRILLO Mauro;
ANGERS Alexandre;
SHIM Jung-Hyun;
TRIJATMIKO Kurniawan;
KREYSA Joachim;
MAZZARA Marco;
2015-02-26
AMER CHEMICAL SOC
JRC91800
0021-8561,
http://pubs.acs.org/doi/abs/10.1021/jf505516y,
https://publications.jrc.ec.europa.eu/repository/handle/JRC91800,
10.1021/jf505516y,
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