Development of Plasmid DNA Reference Material for
the Quantification of Genetically Modified Common Bean Embrapa 5.1
- Publication date
- Publisher
Abstract
The
genetically modified (GM) common bean Embrapa 5.1 was recently
approved for commercialization. The reliable detection and quantification
of GM organisms is strongly dependent on validated methods as well
as calibration systems. This work presents the development of a calibrant
plasmid for Embrapa 5.1 common bean detection. The reaction parameters
were determined and compared for both the plasmid DNA (pDNA) and the
genomic DNA (gDNA). PCR efficiencies for pDNA were 81% for the construction-specific
assays and 76% for the taxon-specific assay, whereas for gDNA efficiencies
were 94 and 93%, respectively. The limits of detection (LOD) in both
qPCR assays were 10<sup>2</sup> and 10<sup>3</sup> copies of gDNA
and pDNA per PCR reaction, respectively. This is sufficient to detect
0.067 and 0.67% of GM common bean in 100 ng of DNA, respectively,
which is in agreement with detecting the 1% GM content required by
the Brazilian legislation