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The utility of optical detection system (qPCR) and bioinformatics methods in reference gene expression analysis

机译:光学检测系统(qPCR)和生物信息学方法在参考基因表达分析中的实用性

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Real-time quantitative polymerase chain reaction is consider as the most reliable method for gene expression studies. However, the expression of target gene could be misinterpreted due to improper normalization. Therefore, the crucial step for analysing of qPCR data is selection of suitable reference genes, which should be validated experimentally. In order to choice the gene with stable expression in the designed experiment, we performed reference gene expression analysis. In this study genes described in the literature and novel genes predicted as control genes, based on the in silico analysis of transcriptome data were used. Analysis with geNorm and NormFinder algorithms allow to create the ranking of candidate genes and indicate the best reference for flower morphogenesis study. According to the results, genes CACS and CYCL were characterised the most stable expression, but the least suitable genes were TUA and EF.
机译:实时定量聚合酶链反应被认为是基因表达研究最可靠的方法。但是,由于归一化不当,可能会误解靶基因的表达。因此,分析qPCR数据的关键步骤是选择合适的参考基因,应通过实验验证。为了在设计的实验中选择表达稳定的基因,我们进行了参考基因表达分析。在这项研究中,使用了基于转录组数据的计算机分析的文献中描述的基因和预测为对照基因的新基因。使用geNorm和NormFinder算法进行的分析可以创建候选基因的排名,并为花卉形态发生研究提供最佳参考。根据结果​​,基因CACS和CYCL被表征为最稳定的表达,而最不适合的基因是TUA和EF。

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