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Int. J. Mol. Sci. 2016, 17(7), 1034;

Evaluation of Housekeeping Genes for Quantitative Real-Time PCR Analysis of Bradysia odoriphaga (Diptera: Sciaridae)

College of Agriculture, Yangtze University, Jingzhou 434025, China
College of Life Science, Heilongjiang University, Harbin 150080, China
Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Department of Medicine, University of Connecticut Health Center, Farmington, CT 06030, USA
These authors contribute equally to this work.
Author to whom correspondence should be addressed.
Academic Editor: Kun Yan Zhu
Received: 8 April 2016 / Revised: 15 June 2016 / Accepted: 20 June 2016 / Published: 7 July 2016
(This article belongs to the Section Biochemistry)
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The soil insect Bradysia odoriphaga (Diptera: Sciaridae) causes substantial damage to Chinese chive. Suitable reference genes in B. odoriphaga (Bradysia odoriphaga) have yet to be identified for normalizing target gene expression among samples by quantitative real-time PCR (qRT-PCR). This study was focused on identifying the expression stability of 12 candidate housekeeping genes in B. odoriphaga under various experiment conditions. The final stability ranking of 12 housekeeping genes was obtained with RefFinder, and the most suitable number of reference genes was analyzed by GeNorm. The results revealed that the most appropriate sets of internal controls were RPS15, RPL18, and RPS18 across developmental phases; RPS15, RPL28, and GAPDH across temperatures; RPS15 and RPL18 across pesticide treatments; RSP5, RPS18, and SDHA across photoperiods; ACTb, RPS18, and RPS15 across diets; RPS13 and RPL28 across populations; and RPS15, ACTb, and RPS18 across all samples. The use of the most suitable reference genes versus an arbitrarily selected reference gene resulted in significant differences in the analysis of a target gene expression. HSP23 in B. odoriphaga was found to be up-regulated under low temperatures. These results will contribute to the standardization of qRT-PCR and will also be valuable for further research on gene function in B. odoriphaga. View Full-Text
Keywords: Bradysia odoriphaga; normalization; reference genes; RefFinder Bradysia odoriphaga; normalization; reference genes; RefFinder

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Shi, C.; Yang, F.; Zhu, X.; Du, E.; Yang, Y.; Wang, S.; Wu, Q.; Zhang, Y. Evaluation of Housekeeping Genes for Quantitative Real-Time PCR Analysis of Bradysia odoriphaga (Diptera: Sciaridae). Int. J. Mol. Sci. 2016, 17, 1034.

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