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Int. J. Mol. Sci. 2014, 15(12), 21825-21839; doi:10.3390/ijms151221825

Real-Time qPCR Identifies Suitable Reference Genes for Borna Disease Virus-Infected Rat Cortical Neurons

1,2,3,†
,
2,3,†
,
2,3,4,†
,
2,3,†
,
5
,
2,3,4
,
2,3
,
2,3
,
2,3
and
1,2,3,4,*
1
Department of Neurology, Yongchuan Hospital, Chongqing Medical University, Chongqing 402460, China
2
Chongqing Key Laboratory of Neurobiology, Chongqing 400016, China
3
Institute of Neuroscience, Chongqing Medical University, Chongqing 400016, China
4
Department of Neurology, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
5
Department of Rehabilitation, the Second Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Received: 14 July 2014 / Revised: 21 September 2014 / Accepted: 25 September 2014 / Published: 26 November 2014
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
View Full-Text   |   Download PDF [903 KB, uploaded 26 November 2014]   |  

Abstract

Quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) is the most commonly-used technique to identify gene expression profiles. The selection of stably expressed reference genes is a prerequisite to properly evaluating gene expression. Here, the suitability of commonly-used reference genes in normalizing RT-qPCR assays of mRNA expression in cultured rat cortical neurons infected with Borna disease virus (BDV) was assessed. The expressions of eight commonly-used reference genes were comparatively analyzed in BDV-infected rat cortical neurons and non-infected control neurons mainly across 9 and 12 days post-infection. These reference genes were validated by RT-qPCR and separately ranked by four statistical algorithms: geNorm, NormFinder, BestKeeper and the comparative delta-Ct method. Then, the RankAggreg package was used to construct consensus rankings. ARBP was found to be the most stable internal control gene at Day 9, and ACTB at Day 12. As the assessment of the validity of the selected reference genes confirms the suitability of applying a combination of the two most stable references genes, combining the two most stable genes for normalization of RT-qPCR studies in BDV-infected rat cortical neurons is recommended at each time point. This study can contribute to improving BDV research by providing the means by which to obtain more reliable and accurate gene expression measurements. View Full-Text
Keywords: Borna disease virus; BDV; reference gene; RT-qPCR; cortical neuron Borna disease virus; BDV; reference gene; RT-qPCR; cortical neuron
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Zhang, L.; Liu, S.; Zhang, L.; You, H.; Huang, R.; Sun, L.; He, P.; Chen, S.; Zhang, H.; Xie, P. Real-Time qPCR Identifies Suitable Reference Genes for Borna Disease Virus-Infected Rat Cortical Neurons. Int. J. Mol. Sci. 2014, 15, 21825-21839.

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