title: Optimization of a multiplex PCR assay for detecting transgenic soybean components in feed products.
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.
authors.affiliation: Key Laboratory of Feed Biotechnology, Ministry of Agriculture, Beijing 100081, People's Republic of China.
authors.forename: Fang
authors.initials: F
authors.lastname: Tian
authors.forename: Xiumin
authors.initials: X
authors.lastname: Wang
authors.forename: Da
authors.initials: D
authors.lastname: Teng
authors.forename: Yalin
authors.initials: Y
authors.lastname: Yang
authors.forename: Qingfeng
authors.initials: Q
authors.lastname: Guan
authors.forename: Changjin
authors.initials: C
authors.lastname: Ao
authors.forename: Jianhua
authors.initials: J
authors.lastname: Wang
mesh_terms: D000821:Animal Feed / Q000032:analysis*; D017931:DNA Primers / Q000235:genetics; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D030821:Plants, Genetically Modified / Q000235:genetics*; D030262:Soybean Proteins / Q000235:genetics; D013025:Glycine max / Q000235:genetics*
chemical_list: D017931:DNA Primers; D030262:Soybean Proteins