Characterization and event-specific quantitative detection of DAS-59122-7 maize insert with the application of plasmidic reference material

BACKGROUND: With the development of genetically modified organisms (GMOs), event-specific qualitative and quantitative polymerase chain reaction (PCR) detection methods have become the internationally agreed standard. RESULTS: The flanking regions of DAS-59122-7 maize were characterized by inverse PCR (I-PCR). In the qualitative PCR assay, a duplex PCR was established with the event-specific and taxon-specific primers, and the limit of detection (LOD) was 1 g kg−1 (approximates to 38 haploid genome copies). In the quantitative TaqMan® real-time PCR assay, a plasmidic reference material was constructed by recombinant PCR and standard curves were set up. By using the plasmidic reference material, we obtained standard curves with good linearity and relatively high efficiency. The results indicated the usability of the plasmid as standard material. CONCLUSION: From above results, we believe that the developed event-specific qualitative and quantitative PCR systems for DAS-59122-7 maize in this study are acceptable and suitable for DAS-59122-7 maize detection. Copyright © 2008 Society of Chemical Industry