中文摘要：开发并商业化地转基因生物数量和多样性的增加促使实验室应用不同的方法对其进行分析。基于矩阵的方法通过减少识别数量来使分析工作最小化，但筛选阶段的正确性需要有有效的方法。本文对来自根癌农杆菌（很多转基因植物中存在的一种遗传成分）的15个胭脂碱合成酶基因终止子（T-NOS）系统进行了测试，T-NOS的引物和探针是通过一种快速、标准的实时PCR技术来合成和测试的。研究表明，快速实时PCR是一种可以在短时间内获得优质数据的可行方法，本研究方法可用于评估筛选寡核苷酸（如引物和探针）。
外文摘要：The increasing number and diversity of genetically modified organisms (GMOs) developed and commercialised forces laboratories to apply many different methods of analysis. Matrix-based approach helps to minimize analytical effort reducing the number of identifications. However, the correctness of screening phase needs efficient methods. In this paper, 15 systems for the nopaline synthase terminator (T-nos) from Agrobacterium tumefaciens, a genetic element present in several genetically modified (GM) plants, were tested. The systems were obtained from three methods, and their primers and probes were combined and tested in real-time polymerase chain reaction (PCR) in fast and standard mode. The results have showed that the fast mode presented the lowest mean quantification cycle (Cq) and the highest a dagger Rn, that is, the difference between normalized reporter and baseline. These parameters of system efficiency prove that the fast real-time PCR is a possible approach to obtain good data in less than half the time compared to standard mode. The proposed approach allows a practical way to evaluate the most efficient set of oligonucleotides (e.g., primers and probe) in fast and standard real-time PCR before validation. This article describes also in-house validation of the best set oligonucleotides.
外文关键词：T-nos(nos-terminator)；Genetically modified organism；Fast real-time PCR；Standard real-time PCR
作者：Elisa Pierboni；Ludovica Curcio；Gloria Raquel Tovo；Martina Torricelli；Cristina Rondini
作者单位：Ist Zooprofilatt Sperimentale Umbria & Marche, Lab OGM & Igiene Ambiente
期刊名称：FOOD ANALYTICAL METHODS
期刊影响因子：2.167
出版年份：2016
出版刊次：4
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