Optimization of a multiplex PCR assay for detecting transgenic soybean components in feed products

文献类型： 外文期刊

第一作者： Wang, Xiumin

作者： Wang, Xiumin;Teng, Da;Yang, Yalin;Guan, Qingfeng;Wang, Jianhua;Tian, Fang;Wang, Xiumin;Teng, Da;Yang, Yalin;Guan, Qingfeng;Wang, Jianhua;Tian, Fang;Ao, Changjin

作者机构：

关键词： Feed;Genetically modified soybean;Multiplex PCR

期刊名称：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY （ 影响因子：2.926； 五年影响因子：2.685 ）

ISSN：

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收录情况： SCI

摘要： 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.

分类号： Q5

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