Direct quantification of picomolar concentrations of mRNAs by mathematical analysis of a reverse transcription/exponential polymerase chain reaction assay.
Direct quantification of picomolar concentrations of mRNAs by mathematical analysis of a reverse transcription/exponential polymerase chain reaction assay.
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通过逆转录/指数聚合酶链反应测定的数学分析直接定量 mRNA 的皮摩尔浓度。
DOI：
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发表时间：
1992
影响因子：
14.9
通讯作者：
Rudolf J. Wiesner
中科院分区：
文献类型：
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作者：
Rudolf J. Wiesner
最近，我们已经证明，通过测量在连续循环中积累的产物的摩尔浓度（n），并分析描述产物积累的方程，log Nn = log effx n + log No，可以准确地确定聚合酶链式反应（PCR）的靶分子数量（No）通过线性回归（1）。在这里，我们将这种方法扩展到通过在扩增之前进行定量逆转录（RT）来测量mRNA的实际拷贝数。由于其高灵敏度，该方法可能成为研究基因表达的有用工具，即使在毫克范围内的样品中，RNA在隔室之间的运输或RNA加工。
Recently, we have shown that the number of target molecules (No) of the polymerase chain reaction (PCR) can be accurately determined by measuring the molar concentration of a product, which accumulates in consecutive cycles (n), and analyzing the equation describing product accumulation, log Nn = log effx n + log No by linear regression (1). Here, we extend this method to the measurement of the actual copy number of mRNAs by performing quantitative reverse transcription (RT) prior to amplification. Due to its high sensitivity, this method might become a useful tool to study gene expression, even in samples in the milligramm range, RNA transport between compartments or RNA processing.