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The focus on sample quality: Influence of colon tissue collection on reliability of qPCR data

机译:关注样品质量:结肠组织收集对qPCR数据可靠性的影响

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摘要

Successful molecular analyses of human solid tissues require intact biological material with well-preserved nucleic acids, proteins, and other cell structures. Pre-analytical handling, comprising of the collection of material at the operating theatre, is among the first critical steps that influence sample quality. The aim of this study was to compare the experimental outcomes obtained from samples collected and stored by the conventional means of snap freezing and by PAXgene Tissue System (Qiagen). These approaches were evaluated by measuring rRNA and mRNA integrity of the samples (RNA Quality Indicator and Differential Amplification Method) and by gene expression profiling. The collection procedures of the biological material were implemented in two hospitals during colon cancer surgery in order to identify the impact of the collection method on the experimental outcome. Our study shows that the pre-analytical sample handling has a significant effect on the quality of RNA and on the variability of qPCR data. PAXgene collection mode proved to be more easily implemented in the operating room and moreover the quality of RNA obtained from human colon tissues by this method is superior to the one obtained by snap freezing.
机译:要对人体实体组织进行成功的分子分析,就需要完整的生物材料以及保存完好的核酸,蛋白质和其他细胞结构。影响样品质量的首要关键步骤之一是进行分析前处理,包括在手术室收集材料。这项研究的目的是比较通过常规速冻和PAXgene Tissue System(Qiagen)从收集和存储的样品中获得的实验结果。通过测量样品的rRNA和mRNA完整性(RNA质量指示剂和差异扩增方法)并通过基因表达谱分析来评估这些方法。为了确定收集方法对实验结果的影响,在结肠癌手术期间在两家医院实施了生物材料的收集程序。我们的研究表明,分析前样品的处理对RNA的质量和qPCR数据的变异性有重大影响。事实证明,PAXgene收集模式在手术室中更容易实施,而且通过这种方法从人结肠组织获得的RNA的质量优于通过速冻获得的RNA的质量。

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