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首页> 外文期刊>Genomics Insights >Genomics Insights: Inter-Laboratory Variability in Array-Based RNA Quantification Methods:
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Genomics Insights: Inter-Laboratory Variability in Array-Based RNA Quantification Methods:

机译：基因组学见解：基于阵列的RNA定量方法的实验室间变异性：

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Ribonucleic acids (RNA) are hypothesized to have preceded their derivatives, deoxyribonucleic acids (DNA), as the molecular media of genetic information when life emerged on earth. Molecular biologists are accustomed to the dramatic effects a subtle variation in the ribose moiety composition between RNA and DNA can have on the stability of these molecules. While DNA is very stable after extraction from biological samples and subsequent treatment, RNA is notoriously labile. The short half-life property, inherent to RNA, benefits cells that do not need to express their entire repertoire of proteins. The cellular machinery turns off the production of a given protein by shutting down the transcription of its cognate coding gene and by either actively degrading the remaining mRNA or allowing it to decay on its own. The steady-state level of each mRNA in a given cell varies continuously and is specified by changing kinetics of synthesis and degradation. Because it is technically possible to simultaneously measure thousands of nucleic acid molecules, these quantities have been studied by the life sciences community to investigate a range of biological problems. Since the RNA abundance can change according to a wide range of perturbations, this makes it the molecule of choice for exploring biological systems; its instability, on the other hand, could be an underestimated source of technical variability. We found that a large fraction of the RNA abundance originally present in the biological system prior to extraction was masked by the RNA labeling and measurement procedure. The method used to extract RNA molecules from cells and to label them prior to hybridization operations on DNA arrays affects the original distribution of RNA. Only if RNA measurements are performed according to the same procedure can biological information be inferred from the assay read out.
机译：据推测，核糖核酸（RNA）在其衍生物脱氧核糖核酸（DNA）之前已经作为地球生命出现时遗传信息的分子介质。分子生物学家习惯于RNA和DNA之间核糖部分组成的细微变化可能对这些分子的稳定性产生巨大影响。从生物样品中提取DNA并进行后续处理后，DNA非常稳定，而RNA非常不稳定。 RNA固有的半衰期短特性使不需要表达其全部蛋白质组成的细胞受益。细胞机制通过关闭其同源编码基因的转录，并通过主动降解剩余的mRNA或使其自身降解来关闭特定蛋白质的产生。给定细胞中每种mRNA的稳态水平连续变化，并通过改变合成和降解动力学来确定。由于在技术上可以同时测量数千个核酸分子，因此生命科学界已经研究了这些数量，以研究一系列生物学问题。由于RNA的丰度会随各种干扰而变化，因此这使其成为探索生物系统的首选分子。另一方面，其不稳定可能是技术可变性的低估来源。我们发现，提取之前最初存在于生物系统中的大部分RNA丰度被RNA标记和测量程序所掩盖。从细胞提取RNA分子并在DNA阵列上进行杂交操作之前对其进行标记的方法会影响RNA的原始分布。只有按照相同的程序进行RNA测量，才能从测定结果中推断出生物学信息。

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