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Selection of Reference Genes in Transcription Analysis of Gene Expression of the Mandarin Fish, Siniperca chuasti.

Research paper by Rui-Xue RX Zhou, Tao T Meng, Hai-Bo HB Meng, Dun-Xue DX Cheng, Shi-Yu SY Bin, Jia J Cheng, Gui-Hong GH Fu, Wu-Ying WY Chu, Jian-She JS Zhang

Indexed on: 15 Jun '10Published on: 15 Jun '10Published in: Dong wu xue yan jiu = Zoological research / "Dong wu xue yan jiu" bian ji wei yuan hui bian ji



Abstract

At present, transcription analysis of gene expression commonly uses housekeeping genes as control for normalization. In this study, the expression levels of three housekeeping genes including GAPDH, beta-actin, and 18S rRNA in six tissues and five developmental stages of the Mandarin fish Siniperca chuatsi were assayed with quantitative real-time PCR (qPCR). Differences in expression levels were analyzed using geNorm program. The results demonstrate that beta-actin is the most stable gene at developmental stages and GAPDH is the most stable in different tissues. While 18S rRNA expression during development is differentially regulated, which indicates it is suitable as an internal control for gene expression normalization at the developmental level. Overall, the data suggest that the two most stable housekeeping genes are enough to accurately calibrate gene expression in S. chuatsi. The significance of this study provided convincing references and methodology for housekeeping gene selection and normalization in gene expression analysis with regular PCR or qPCR.