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EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR
Published on: January 16, 2016
Validation of Stable Reference Genes for RT-qPCR Normalization in Oxycetonia jucunda (Coleoptera: Scarabaeidae)
Shi-Hang Zhao1,2, Yang Yue3, Rui-Tao Yu1,2
1Shijiazhuang Institute of Pomology, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050061, China.
Abstract:
The polyphagous pest Oxycetonia jucunda Faldermann can cause substantial damage to a range of economically important crops, with the adult beetles feeding directly on floral tissues and young leaves. RT-qPCR is widely used to analyze gene expression, for which the selection of stable reference genes is essential for enabling an accurate normalization of expression. However, no systematic evaluations of suitable reference genes for RT-qPCR analysis using different tissues of O. jucunda have been conducted. To assess their applicability as reliable normalization controls, we used five computational methods to examine the stability of seven potential reference genes (GAPDH, EF1α, RPS3, RPS18, RPL18, RPS31, and UBC5A) across six adult tissues, with three biological replicates per tissue. The findings revealed RPS3 and RPS31 to be the most stably expressed. This pair of reference genes was further validated by normalizing the expression of the odorant-binding protein 3 (OBP3) target gene. Our findings will provide important foundational data for the accurate analysis of functional gene expression in O. jucunda.
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