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Reference Gene Selection for qPCR Normalization of Kosteletzkya virginica under Salt Stress

Kosteletzkya virginica (L.) is a newly introduced perennial halophytic plant. Presently, reverse transcription quantitative real-time PCR (qPCR) is regarded as the best choice for analyzing gene expression and its accuracy mainly depends on the reference genes which are used for gene expression norm...

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Detalles Bibliográficos
Autores principales: Tang, Xiaoli, Wang, Hongyan, Shao, Chuyang, Shao, Hongbo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4637064/
https://www.ncbi.nlm.nih.gov/pubmed/26581422
http://dx.doi.org/10.1155/2015/823806
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author Tang, Xiaoli
Wang, Hongyan
Shao, Chuyang
Shao, Hongbo
author_facet Tang, Xiaoli
Wang, Hongyan
Shao, Chuyang
Shao, Hongbo
author_sort Tang, Xiaoli
collection PubMed
description Kosteletzkya virginica (L.) is a newly introduced perennial halophytic plant. Presently, reverse transcription quantitative real-time PCR (qPCR) is regarded as the best choice for analyzing gene expression and its accuracy mainly depends on the reference genes which are used for gene expression normalization. In this study, we employed qPCR to select the most stable reference gene in K. virginica which showed stable expression profiles under our experimental conditions. The candidate reference genes were 18S ribosomal RNA (18SrRNA), β-actin (ACT), α-tubulin (TUA), and elongation factor (EF). We tracked the gene expression profiles of the candidate genes and analyzed their stabilities through BestKeeper, geNorm, and NormFinder software programs. The results of the three programs were identical and 18SrRNA was assessed to be the most stable reference gene in this study. However, TUA was identified to be the most unstable. Our study proved again that the traditional reference genes indeed displayed a certain degree of variations under given experimental conditions. Importantly, our research also provides guidance for selecting most suitable reference genes and lays the foundation for further studies in K. virginica.
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spelling pubmed-46370642015-11-18 Reference Gene Selection for qPCR Normalization of Kosteletzkya virginica under Salt Stress Tang, Xiaoli Wang, Hongyan Shao, Chuyang Shao, Hongbo Biomed Res Int Research Article Kosteletzkya virginica (L.) is a newly introduced perennial halophytic plant. Presently, reverse transcription quantitative real-time PCR (qPCR) is regarded as the best choice for analyzing gene expression and its accuracy mainly depends on the reference genes which are used for gene expression normalization. In this study, we employed qPCR to select the most stable reference gene in K. virginica which showed stable expression profiles under our experimental conditions. The candidate reference genes were 18S ribosomal RNA (18SrRNA), β-actin (ACT), α-tubulin (TUA), and elongation factor (EF). We tracked the gene expression profiles of the candidate genes and analyzed their stabilities through BestKeeper, geNorm, and NormFinder software programs. The results of the three programs were identical and 18SrRNA was assessed to be the most stable reference gene in this study. However, TUA was identified to be the most unstable. Our study proved again that the traditional reference genes indeed displayed a certain degree of variations under given experimental conditions. Importantly, our research also provides guidance for selecting most suitable reference genes and lays the foundation for further studies in K. virginica. Hindawi Publishing Corporation 2015 2015-10-25 /pmc/articles/PMC4637064/ /pubmed/26581422 http://dx.doi.org/10.1155/2015/823806 Text en Copyright © 2015 Xiaoli Tang et al. https://creativecommons.org/licenses/by/3.0/ 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.
spellingShingle Research Article
Tang, Xiaoli
Wang, Hongyan
Shao, Chuyang
Shao, Hongbo
Reference Gene Selection for qPCR Normalization of Kosteletzkya virginica under Salt Stress
title Reference Gene Selection for qPCR Normalization of Kosteletzkya virginica under Salt Stress
title_full Reference Gene Selection for qPCR Normalization of Kosteletzkya virginica under Salt Stress
title_fullStr Reference Gene Selection for qPCR Normalization of Kosteletzkya virginica under Salt Stress
title_full_unstemmed Reference Gene Selection for qPCR Normalization of Kosteletzkya virginica under Salt Stress
title_short Reference Gene Selection for qPCR Normalization of Kosteletzkya virginica under Salt Stress
title_sort reference gene selection for qpcr normalization of kosteletzkya virginica under salt stress
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4637064/
https://www.ncbi.nlm.nih.gov/pubmed/26581422
http://dx.doi.org/10.1155/2015/823806
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