Validation of reference genes for use in untreated bovine fibroblasts

Abstract Proper normalization of RT-qPCR data is pivotal to the interpretation of results and accuracy of scientific conclusions. Though different approaches may be taken, normalization against multiple reference genes is now standard practice. Genes traditionally used and deemed constitutively expr...

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Autores principales:	T. Toorani, P. M. Mackie, G. F. Mastromonaco
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/71772de2425543b0be0f4a4216fe9a5b
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spelling	oai:doaj.org-article:71772de2425543b0be0f4a4216fe9a5b2021-12-02T16:50:17ZValidation of reference genes for use in untreated bovine fibroblasts10.1038/s41598-021-89657-82045-2322https://doaj.org/article/71772de2425543b0be0f4a4216fe9a5b2021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-89657-8https://doaj.org/toc/2045-2322Abstract Proper normalization of RT-qPCR data is pivotal to the interpretation of results and accuracy of scientific conclusions. Though different approaches may be taken, normalization against multiple reference genes is now standard practice. Genes traditionally used and deemed constitutively expressed have demonstrated variability in expression under different experimental conditions, necessitating the proper validation of reference genes prior to utilization. Considering the wide use of fibroblasts in research and scientific applications, it is imperative that suitable reference genes for fibroblasts of different animal origins and conditions be elucidated. Previous studies on bovine fibroblasts have tested limited genes and/or samples. Herein, we present an extensive study investigating the expression stability of 16 candidate reference genes across 7 untreated bovine fibroblast cell lines subjected to controlled conditions. Data were analysed using various statistical tools and algorithms, including geNorm, NormFinder, BestKeeper, and RefFinder. A combined use of GUSB and RPL13A was determined to be the best approach for data normalization in untreated bovine fibroblasts.T. TooraniP. M. MackieG. F. MastromonacoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q T. Toorani P. M. Mackie G. F. Mastromonaco Validation of reference genes for use in untreated bovine fibroblasts
description	Abstract Proper normalization of RT-qPCR data is pivotal to the interpretation of results and accuracy of scientific conclusions. Though different approaches may be taken, normalization against multiple reference genes is now standard practice. Genes traditionally used and deemed constitutively expressed have demonstrated variability in expression under different experimental conditions, necessitating the proper validation of reference genes prior to utilization. Considering the wide use of fibroblasts in research and scientific applications, it is imperative that suitable reference genes for fibroblasts of different animal origins and conditions be elucidated. Previous studies on bovine fibroblasts have tested limited genes and/or samples. Herein, we present an extensive study investigating the expression stability of 16 candidate reference genes across 7 untreated bovine fibroblast cell lines subjected to controlled conditions. Data were analysed using various statistical tools and algorithms, including geNorm, NormFinder, BestKeeper, and RefFinder. A combined use of GUSB and RPL13A was determined to be the best approach for data normalization in untreated bovine fibroblasts.
format	article
author	T. Toorani P. M. Mackie G. F. Mastromonaco
author_facet	T. Toorani P. M. Mackie G. F. Mastromonaco
author_sort	T. Toorani
title	Validation of reference genes for use in untreated bovine fibroblasts
title_short	Validation of reference genes for use in untreated bovine fibroblasts
title_full	Validation of reference genes for use in untreated bovine fibroblasts
title_fullStr	Validation of reference genes for use in untreated bovine fibroblasts
title_full_unstemmed	Validation of reference genes for use in untreated bovine fibroblasts
title_sort	validation of reference genes for use in untreated bovine fibroblasts
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/71772de2425543b0be0f4a4216fe9a5b
work_keys_str_mv	AT ttoorani validationofreferencegenesforuseinuntreatedbovinefibroblasts AT pmmackie validationofreferencegenesforuseinuntreatedbovinefibroblasts AT gfmastromonaco validationofreferencegenesforuseinuntreatedbovinefibroblasts
_version_	1718383044725833728

Validation of reference genes for use in untreated bovine fibroblasts

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