Validation of reference genes for quantitative real-time PCR in tiger beetles across sexes, body parts, sexual maturity and immune challenge

Abstract Reference genes are frequently used as normalizers for expression studies despite not being previously verified to present suitable stabilities. Considering the interest that tiger beetles have generated in the past years, resulting in a variety of studies, it is crucial to dispose of a val...

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Autores principales:	Andrés García-Reina, María Juliana Rodríguez-García, José Galián
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/45eca28f959543519aa74b2b7c3eceb3
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spelling	oai:doaj.org-article:45eca28f959543519aa74b2b7c3eceb32021-12-02T12:32:48ZValidation of reference genes for quantitative real-time PCR in tiger beetles across sexes, body parts, sexual maturity and immune challenge10.1038/s41598-018-28978-72045-2322https://doaj.org/article/45eca28f959543519aa74b2b7c3eceb32018-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-28978-7https://doaj.org/toc/2045-2322Abstract Reference genes are frequently used as normalizers for expression studies despite not being previously verified to present suitable stabilities. Considering the interest that tiger beetles have generated in the past years, resulting in a variety of studies, it is crucial to dispose of a validated reference gene panel for expression studies. Nine candidate genes were tested in Cicindela campestris and Calomera littoralis across several conditions and their transcription levels were assessed with geNorm, NormFinder, BestKeeper and ΔC T method algorithms. Results showed high stabilities across sexes, immune challenge and gonad developmental stages for all genes tested, while body parts comparison presented less constant expression values. Only two genes are sufficient to perform a proper normalization for most of the conditions tested, except for the body parts comparison in C. littoralis, which requires the use of at least three reference genes. On the whole, no universal gene is found to be suitable for all situations, but according to the acceptable range of values, NADH, B-t, Vatpase and ArgKin seem to present the most constant expression stability, indicating their suitability as reference genes in most of the conditions. This is the first report evaluating the stability of housekeeping genes in adephagan beetles.Andrés García-ReinaMaría Juliana Rodríguez-GarcíaJosé GaliánNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-13 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Andrés García-Reina María Juliana Rodríguez-García José Galián Validation of reference genes for quantitative real-time PCR in tiger beetles across sexes, body parts, sexual maturity and immune challenge
description	Abstract Reference genes are frequently used as normalizers for expression studies despite not being previously verified to present suitable stabilities. Considering the interest that tiger beetles have generated in the past years, resulting in a variety of studies, it is crucial to dispose of a validated reference gene panel for expression studies. Nine candidate genes were tested in Cicindela campestris and Calomera littoralis across several conditions and their transcription levels were assessed with geNorm, NormFinder, BestKeeper and ΔC T method algorithms. Results showed high stabilities across sexes, immune challenge and gonad developmental stages for all genes tested, while body parts comparison presented less constant expression values. Only two genes are sufficient to perform a proper normalization for most of the conditions tested, except for the body parts comparison in C. littoralis, which requires the use of at least three reference genes. On the whole, no universal gene is found to be suitable for all situations, but according to the acceptable range of values, NADH, B-t, Vatpase and ArgKin seem to present the most constant expression stability, indicating their suitability as reference genes in most of the conditions. This is the first report evaluating the stability of housekeeping genes in adephagan beetles.
format	article
author	Andrés García-Reina María Juliana Rodríguez-García José Galián
author_facet	Andrés García-Reina María Juliana Rodríguez-García José Galián
author_sort	Andrés García-Reina
title	Validation of reference genes for quantitative real-time PCR in tiger beetles across sexes, body parts, sexual maturity and immune challenge
title_short	Validation of reference genes for quantitative real-time PCR in tiger beetles across sexes, body parts, sexual maturity and immune challenge
title_full	Validation of reference genes for quantitative real-time PCR in tiger beetles across sexes, body parts, sexual maturity and immune challenge
title_fullStr	Validation of reference genes for quantitative real-time PCR in tiger beetles across sexes, body parts, sexual maturity and immune challenge
title_full_unstemmed	Validation of reference genes for quantitative real-time PCR in tiger beetles across sexes, body parts, sexual maturity and immune challenge
title_sort	validation of reference genes for quantitative real-time pcr in tiger beetles across sexes, body parts, sexual maturity and immune challenge
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/45eca28f959543519aa74b2b7c3eceb3
work_keys_str_mv	AT andresgarciareina validationofreferencegenesforquantitativerealtimepcrintigerbeetlesacrosssexesbodypartssexualmaturityandimmunechallenge AT mariajulianarodriguezgarcia validationofreferencegenesforquantitativerealtimepcrintigerbeetlesacrosssexesbodypartssexualmaturityandimmunechallenge AT josegalian validationofreferencegenesforquantitativerealtimepcrintigerbeetlesacrosssexesbodypartssexualmaturityandimmunechallenge
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Validation of reference genes for quantitative real-time PCR in tiger beetles across sexes, body parts, sexual maturity and immune challenge

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