title: High-throughput, low-cost, and event-specific polymerase chain reaction detection of herbicide tolerance in genetically modified soybean A2704-12.
abstract: The aim of this study was to develop an event-specific qualitative and real-time quantitative polymerase chain reaction (PCR) method for detection of herbicide-tolerance genetically modified (GM) soybean A2704-12. The event-specific PCR primers were designed, based on the 5'-flanking integration sequence in the soybean genome, to amplify the 239-bp target fragment. Employing the same event-specific primers, qualitative PCR and real-time quantitative PCR detection methods were successfully developed. The results showed that the A2704-12 event could be specifically distinguished from other GM soybean events. In the qualitative PCR assay, the limit of detection was 0.05%, and in the real-time quantitative PCR assay, the limit of detection was less than 0.01%. Moreover, our genomic DNA (gDNA) extraction protocol is high-throughput, safe, and low-cost. The event-specific PCR assay system is cost-efficient by using SYBR Green I in real-time PCR, and by using the same primers in both the qualitative and quantitative PCR assays. We therefore developed a high-throughput, low-cost, and event-specific qualitative and quantitative PCR detection method for GM soybean A2704-12. The method would be useful for market supervision and management of GM soybean A2704-12 due to its high specificity and sensitivity.
authors.affiliation: College of Life Sciences, Anhui Agricultural University, Hefei, China.
authors.forename: H
authors.initials: H
authors.lastname: Ma
authors.affiliation: GMOs and Derived Products Inspection and Supervision Center (Hefei), Ministry of Agriculture/Key Laboratory of Rice Genetics Breeding of Anhui Province/Rice Research Institute, Anhui Academy of Agricultural Sciences, Hefei, China.
authors.forename: H
authors.initials: H
authors.lastname: Li
authors.affiliation: GMOs and Derived Products Inspection and Supervision Center (Hefei), Ministry of Agriculture/Key Laboratory of Rice Genetics Breeding of Anhui Province/Rice Research Institute, Anhui Academy of Agricultural Sciences, Hefei, China.
authors.forename: J
authors.initials: J
authors.lastname: Li
authors.affiliation: GMOs and Derived Products Inspection and Supervision Center (Hefei), Ministry of Agriculture/Key Laboratory of Rice Genetics Breeding of Anhui Province/Rice Research Institute, Anhui Academy of Agricultural Sciences, Hefei, China.
authors.forename: X F
authors.initials: XF
authors.lastname: Wang
authors.affiliation: GMOs and Derived Products Inspection and Supervision Center (Hefei), Ministry of Agriculture/Key Laboratory of Rice Genetics Breeding of Anhui Province/Rice Research Institute, Anhui Academy of Agricultural Sciences, Hefei, China.
authors.forename: P C
authors.initials: PC
authors.lastname: Wei
authors.affiliation: GMOs and Derived Products Inspection and Supervision Center (Hefei), Ministry of Agriculture/Key Laboratory of Rice Genetics Breeding of Anhui Province/Rice Research Institute, Anhui Academy of Agricultural Sciences, Hefei, China.
authors.forename: L
authors.initials: L
authors.lastname: Li
authors.affiliation: GMOs and Derived Products Inspection and Supervision Center (Hefei), Ministry of Agriculture/Key Laboratory of Rice Genetics Breeding of Anhui Province/Rice Research Institute, Anhui Academy of Agricultural Sciences, Hefei, China yjianbo@263.net.
authors.forename: J B
authors.initials: JB
authors.lastname: Yang
mesh_terms: D001483:Base Sequence; D006540:Herbicides / Q000633:toxicity; D057166:High-Throughput Screening Assays / Q000191:economics / Q000379:methods*; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics / Q000254:growth & development; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D013025:Glycine max / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development
chemical_list: D006540:Herbicides