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CURVE PROCESSOR ALGORITHM FOR THE QUALITY CONTROL OF (RT-)qPCR CURVES

机译:(RT-)qPCR曲线质量控制的曲线处理器算法

摘要

The invention is in the field of analytical technology and relates to an improved procedure for determining the concentration or activity of an analyte in a sample. Specifically the invention provides an automated algorithm for the quality control of (RT-)qPCR reactions. Plotting the fluorescence intensity of a reporter dye divided by the fluorescence intensity of a passive reference dye against the cycle number leads to a so-called sigmoid function which is characterized by a background phase, an exponential growth phase and a plateau phase. Since the fluorescence intensity as a function of cycles relates to the initial number of template molecules in the sample, qPCR curves can be used to quantify the amount of RNA or DNA fragments in the sample by determination of a so-called Cq value.
机译:本发明属于分析技术领域,并且涉及一种用于确定样品中分析物的浓度或活性的改进方法。具体地,本发明提供了用于(RT-)qPCR反应的质量控制的自动化算法。将报告染料的荧光强度除以被动参比染料的荧光强度相对于循环数作图可得出所谓的S形函数,其特征在于背景阶段,指数生长期和平稳阶段。由于荧光强度是循环的函数,与样品中模板分子的初始数量有关,因此qPCR曲线可用于通过确定所谓的Cq值来量化样品中RNA或DNA片段的数量。

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