Selection of <i>Bactrocera tau</i> (Walker) Reference Genes for Quantitative Real-Time PCR

The selection of appropriate reference genes is critical for standardizing quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) data, thereby ensuring accurate and reliable results of gene expression analysis. In this study, we identified 10 candidate reference genes (enc...

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Bibliographic Details
Main Authors: Yutong Zhai, Yonghao Yu, Pengfei Xu, Xianru Zeng, Xiuzhen Long, Dewei Wei, Zhan He, Xuyuan Gao
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Insects
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Online Access:https://www.mdpi.com/2075-4450/16/5/445
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Summary:The selection of appropriate reference genes is critical for standardizing quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) data, thereby ensuring accurate and reliable results of gene expression analysis. In this study, we identified 10 candidate reference genes (encoding <i>α-tubulin</i>, <i>G6PDH</i>, <i>Rab1</i>, <i>RT</i>, <i>RPS13</i>, <i>β-tubulin</i>, <i>DPH1</i>, <i>HSP90</i>, <i>GAPDH</i>, and <i>CP</i>) and evaluated their suitability for use as reference genes in the pest insect, <i>Bactrocera tau</i>. Analysis was conducted using three software-based methods—Delta CT, NormFinder, and BestKeeper—alongside the online tool RefFinder. Expression levels of these genes were analyzed across various <i>B. tau</i> developmental stages and body parts. The overall ranking of reference gene stability scores was as follows: <i>α-tubulin</i> > <i>G6PDH</i> > <i>CP</i> > <i>β-tubulin</i> > <i>RT</i> > <i>HSP90</i> > <i>GAPDH</i> > <i>DPH1</i> > <i>RPS13</i> > <i>Rab1</i>. Ultimately, <i>α-tubulin</i> and <i>G6PDH</i> were identified as the most stable reference genes for <i>B. tau</i>.
ISSN:2075-4450