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Selection of a suitable reference gene for quantitative gene expression in mouse lymph nodes after vaccination

BACKGROUND: The quantification of gene expression is an important tool in the evaluation of the immune response to vaccines. Reliable reference genes for gene expression studies in mouse draining lymph nodes after vaccination have not been reported. RESULTS: The utility of seven potential reference...

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Autores principales: Mosley, Yung-Yi C., HogenEsch, Harm
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5718095/
https://www.ncbi.nlm.nih.gov/pubmed/29208024
http://dx.doi.org/10.1186/s13104-017-3005-y
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author Mosley, Yung-Yi C.
HogenEsch, Harm
author_facet Mosley, Yung-Yi C.
HogenEsch, Harm
author_sort Mosley, Yung-Yi C.
collection PubMed
description BACKGROUND: The quantification of gene expression is an important tool in the evaluation of the immune response to vaccines. Reliable reference genes for gene expression studies in mouse draining lymph nodes after vaccination have not been reported. RESULTS: The utility of seven potential reference genes was investigated using commercially available Taq-man primer/probe mixes. Results were evaluated with RefFinder, a web-based program including multiple algorithm methods such as geNorm, NormFinder, BestKeeper and the comparative delta-Ct. Further assessment was done by applying the candidate reference genes in relative expression calculations with genes related to the magnitude and longevity of the humoral immune responses. The ubiquitin C gene, Ubc, was found to be the most reliable reference gene when validated with well-known genes that are expressed at relatively low levels after vaccination. The optimal time of sample collection varied depending on the function of the target genes. CONCLUSIONS: This study identified Ubc as the most reliable reference gene and provides useful information for studies examining immunological gene expression in the draining lymph nodes after vaccination in mice. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13104-017-3005-y) contains supplementary material, which is available to authorized users.
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spelling pubmed-57180952017-12-08 Selection of a suitable reference gene for quantitative gene expression in mouse lymph nodes after vaccination Mosley, Yung-Yi C. HogenEsch, Harm BMC Res Notes Research Article BACKGROUND: The quantification of gene expression is an important tool in the evaluation of the immune response to vaccines. Reliable reference genes for gene expression studies in mouse draining lymph nodes after vaccination have not been reported. RESULTS: The utility of seven potential reference genes was investigated using commercially available Taq-man primer/probe mixes. Results were evaluated with RefFinder, a web-based program including multiple algorithm methods such as geNorm, NormFinder, BestKeeper and the comparative delta-Ct. Further assessment was done by applying the candidate reference genes in relative expression calculations with genes related to the magnitude and longevity of the humoral immune responses. The ubiquitin C gene, Ubc, was found to be the most reliable reference gene when validated with well-known genes that are expressed at relatively low levels after vaccination. The optimal time of sample collection varied depending on the function of the target genes. CONCLUSIONS: This study identified Ubc as the most reliable reference gene and provides useful information for studies examining immunological gene expression in the draining lymph nodes after vaccination in mice. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13104-017-3005-y) contains supplementary material, which is available to authorized users. BioMed Central 2017-12-06 /pmc/articles/PMC5718095/ /pubmed/29208024 http://dx.doi.org/10.1186/s13104-017-3005-y Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Mosley, Yung-Yi C.
HogenEsch, Harm
Selection of a suitable reference gene for quantitative gene expression in mouse lymph nodes after vaccination
title Selection of a suitable reference gene for quantitative gene expression in mouse lymph nodes after vaccination
title_full Selection of a suitable reference gene for quantitative gene expression in mouse lymph nodes after vaccination
title_fullStr Selection of a suitable reference gene for quantitative gene expression in mouse lymph nodes after vaccination
title_full_unstemmed Selection of a suitable reference gene for quantitative gene expression in mouse lymph nodes after vaccination
title_short Selection of a suitable reference gene for quantitative gene expression in mouse lymph nodes after vaccination
title_sort selection of a suitable reference gene for quantitative gene expression in mouse lymph nodes after vaccination
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5718095/
https://www.ncbi.nlm.nih.gov/pubmed/29208024
http://dx.doi.org/10.1186/s13104-017-3005-y
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