文章基本信息

标题：Validation of house-keeping genes for normalization of gene expression data during diurnal/circadian studies in rice by RT-qPCR
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作者：Nitin Jain ; Satyam Vergish ; Jitendra P. Khurana 等
期刊名称：Scientific Reports
电子版ISSN：2045-2322
出版年度：2018
卷号：8
期号：1
页码：3203
DOI：10.1038/s41598-018-21374-1
语种：English
出版社：Springer Nature
摘要：The circadian clock in plants is the intrinsic rhythmic expression of thousands of genes in a 24 h period, which is set by the day-night cycles in the environment. The study of the circadian clock often requires expression profiling of genes over a large number of samples for which RT-qPCR is invariably used. Reliability of the results depends largely on the house-keeping genes, which serve as control and thus should be chosen carefully to prevent erroneous results. In this study, ten house-keeping genes were chosen from rice for stability analysis with 48 tissue samples harvested from plants subjected to diurnal/circadian cycles. Although, all the genes were found to be stable, however, six of them showed cyclic expression patterns and caused major changes in the expression profiles of the target genes when used to normalize their expression data, thereby making them poor candidates for diurnal/circadian studies. In conclusion, reference genes need to be selected for diurnal/circadian studies with extra caution as more than 80% of transcriptome in plants undergoes cycling, which remains undetected by the gene stability assessment software and can severely affect the RT-qPCR based gene expression profiling. The geometric mean of two or more most stable reference genes is hence recommended for diurnal/circadian studies in plants.