Development of the One-step Visual Loop-Mediated Isothermal Amplification Assay for Genetically Modified Rice Event TT51-1
文献类型： 外文期刊
作者： Chen, Rui 1 ; Wang, Yong 1 ; Zhu, Zhu 1 ; Lan, Qingkuo 1 ; Zhao, Xin 1 ; Wang, Haiyang 3 ; Guo, Yongze 1 ; Wang, Qinying 2 ;
作者机构： 1.Tianjin Acad Agr Sci, Tianjin Inst Agr Qual Stand & Testing Technol, Tianjin 300192, Peoples R China
2.Hebei Agr Univ, Coll Plant Protect, Baoding 071001, Peoples R China
3.Qinhuangdao Entry Exit Inspect & Quarantine Bur, Qinhuangdao 066004, Peoples R China
关键词： loop-mediated isothermal amplification;genetically modified crop;rice;TT51-1
期刊名称：FOOD SCIENCE AND TECHNOLOGY RESEARCH （ 影响因子：0.668； 五年影响因子：0.895 ）
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收录情况： SCI
摘要： Loop-mediated isothermal amplification (LAMP), a novel DNA amplification technique, is widely used in molecular diagnosis although it is very easy to be contaminated by the huge amount of its own amplicons. An improved visual LAMP method for the event specific identification of GM rice TT51-1 and a rice endogenous reference gene (sucrose phosphate synthase, SPS) was established. Pre-loaded calcein and manganous ion into the LAMP reaction offered not only an easy way to read results by colour change from orange to green but also a closed-tube system to reduce contaminations caused by amplification products. The LAMP assays could be finished within 60 min, and the limits of detection (LODs) were estimated as low as 10 and 20 copies of rice haploid genomic DNA, respectively, about 10-fold more sensitive than that of conventional PCR. The specificities of the assays were also well evaluated with optimized system and conditions. The developed one-step visual LAMP assays in this study provided a promising tool for the convenient and effective detection of GM crops.
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