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Identification and validation of Aeluropus littoralis reference genes for Quantitative Real-Time PCR Normalization

机译:定量和实时定量PCR标准化的滨海臭参参考基因的鉴定和验证

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摘要

BackgroundThe use of stably expressed genes as normalizers has crucial role in accurate and reliable expression analysis estimated by quantitative real-time polymerase chain reaction (qPCR). Recent studies have shown that, the expression levels of common housekeeping genes are varying in different tissues and experimental conditions. The genomic DNA contamination in RNA samples is another important factor that also influence the interpretation of the data obtained from qPCR. It is estimated that the gDNA contamination in gene expression analysis lead to an overestimation of the RNA transcript level. The aim of this study was to validate the most stably expressed reference genes in two different tissues of Aeluropus littoralis—halophyte grass at salt stress and recovery condition. Also, a qPCR-based approach for monitoring contamination with gDNA was conducted.
机译:背景技术使用稳定表达的基因作为标准化试剂在通过定量实时聚合酶链反应(qPCR)进行的准确可靠的表达分析中起着至关重要的作用。最近的研究表明,普通管家基因的表达水平在不同的组织和实验条件下是不同的。 RNA样品中的基因组DNA污染是另一个重要因素,它也影响从qPCR获得的数据的解释。据估计,基因表达分析中的gDNA污染会导致RNA转录水平的高估。这项研究的目的是验证盐胁迫和恢复条件下,东北拟南芥(Aeluropus littoralis)-盐生植物的两个不同组织中表达最稳定的参考基因。此外,还进行了基于qPCR的方法监测gDNA污染。

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