Abstract
A multiplex polymerase chain reaction (m-PCR) assay was developed for the simultaneous detection of multiple components of genetically modified (GM) soybean. It uses two sets of primers (I, lectin1/35S/CP4; II, lectin2/35S/CP4) specific for a soybean reference gene, the 35S promoter, and an event-specific gene. Amplified fragments of 118, 414, 195, and 320 bp were easily detected by agarose gel electrophoresis and were positively confirmed by sequencing. Primer set concentrations and annealing temperatures in the m-PCR were optimized. The optimized m-PCR conditions were obtained for primer set I at a ratio of 1:2:3 and a 59.2 °C annealing temperature and set II at the same ratio and 58.6 °C, 60.3 °C, and 61.2 °C annealing temperatures. The sensitivities of the two m-PCR primer sets (I and II) were 0.25% and 0.5%, respectively. The results showed that this m-PCR assay provides rapid, reliable, and effective identification of multiple components of GM soybean in feed.
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- GM:
-
Genetically modified
- RRS:
-
Roundup ready soybean
- CRMs:
-
Certified reference materials
- m-PCR:
-
Multiplex PCR
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Acknowledgments
This study is supported by National Major Project of Breeding for Genetically Modified Organism in China (no. 2009ZX08012-012B).
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Fang Tian and Xiumin Wang contributed equally to this paper.
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Tian, F., Wang, X., Teng, D. et al. Optimization of a Multiplex PCR Assay for Detecting Transgenic Soybean Components in Feed Products. Appl Biochem Biotechnol 165, 1225–1234 (2011). https://doi.org/10.1007/s12010-011-9340-x
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DOI: https://doi.org/10.1007/s12010-011-9340-x