Analysis of qPCR reference genes stability determination methods and a practical approach for efficiency calculation on a turbot (Scphthalmus maximus) gonad dataset
Gene expression analysis by reverse transcription quantitative PCR (qPCR) is the most widely used method for analyzing the expression of a moderate number of genes and also for the validation of microarray results. Several issues are crucial for a successful qPCR study, particularly the selection of...
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Online Access: | http://hdl.handle.net/10508/2045 https://doi.org/10.1186/1471-2164-15-648 |
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ftieo:oai:repositorio.ieo.es:10508/2045 2023-05-15T18:15:43+02:00 Analysis of qPCR reference genes stability determination methods and a practical approach for efficiency calculation on a turbot (Scphthalmus maximus) gonad dataset Robledo, D. (Diego) Hernández-Urcera, J. (Jorge) Cal, R. (Rosa) Pardo, B. (Belén) Sánchez, L. (Laura) Martínez, P. (Paulino) Viñas, A. (Ana) 2014-08-04 http://hdl.handle.net/10508/2045 https://doi.org/10.1186/1471-2164-15-648 eng eng Centro Oceanográfico de Vigo http://hdl.handle.net/10508/2045 BMC Genomics, 15. 2014: 648-657 doi:10.1186/1471-2164-15-648 Atribución-NoComercial-SinDerivadas 3.0 España http://creativecommons.org/licenses/by-nc-nd/3.0/es/ restrictedAccess CC-BY-NC-ND qPCR Reference genes Amplification efficiency Turbot Scophthalmus maximus article 2014 ftieo https://doi.org/10.1186/1471-2164-15-648 2022-08-30T23:46:46Z Gene expression analysis by reverse transcription quantitative PCR (qPCR) is the most widely used method for analyzing the expression of a moderate number of genes and also for the validation of microarray results. Several issues are crucial for a successful qPCR study, particularly the selection of internal reference genes for normalization and efficiency determination. There is no agreement on which method is the best to detect the most stable genes neither on how to perform efficiency determination. In this study we offer a comprehensive evaluation of the characteristics of reference gene selection methods and how to decide which one is more reliable when they show discordant outcomes. Also, we analyze the current efficiency calculation controversy. Our dataset is composed by gonad samples of turbot at different development times reared at different temperatures. Turbot (Scophthalmus maximus) is a relevant marine aquaculture European species with increasing production in the incoming years. Since females largely outgrow males, identification of genes related to sex determination, gonad development and reproductive behavior, and analysis of their expression profiles are of primary importance for turbot industry Versión del editor Article in Journal/Newspaper Scophthalmus maximus Turbot Instituto Español de Oceanografía: e-IEO BMC Genomics 15 1 648 |
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Open Polar |
collection |
Instituto Español de Oceanografía: e-IEO |
op_collection_id |
ftieo |
language |
English |
topic |
qPCR Reference genes Amplification efficiency Turbot Scophthalmus maximus |
spellingShingle |
qPCR Reference genes Amplification efficiency Turbot Scophthalmus maximus Robledo, D. (Diego) Hernández-Urcera, J. (Jorge) Cal, R. (Rosa) Pardo, B. (Belén) Sánchez, L. (Laura) Martínez, P. (Paulino) Viñas, A. (Ana) Analysis of qPCR reference genes stability determination methods and a practical approach for efficiency calculation on a turbot (Scphthalmus maximus) gonad dataset |
topic_facet |
qPCR Reference genes Amplification efficiency Turbot Scophthalmus maximus |
description |
Gene expression analysis by reverse transcription quantitative PCR (qPCR) is the most widely used method for analyzing the expression of a moderate number of genes and also for the validation of microarray results. Several issues are crucial for a successful qPCR study, particularly the selection of internal reference genes for normalization and efficiency determination. There is no agreement on which method is the best to detect the most stable genes neither on how to perform efficiency determination. In this study we offer a comprehensive evaluation of the characteristics of reference gene selection methods and how to decide which one is more reliable when they show discordant outcomes. Also, we analyze the current efficiency calculation controversy. Our dataset is composed by gonad samples of turbot at different development times reared at different temperatures. Turbot (Scophthalmus maximus) is a relevant marine aquaculture European species with increasing production in the incoming years. Since females largely outgrow males, identification of genes related to sex determination, gonad development and reproductive behavior, and analysis of their expression profiles are of primary importance for turbot industry Versión del editor |
format |
Article in Journal/Newspaper |
author |
Robledo, D. (Diego) Hernández-Urcera, J. (Jorge) Cal, R. (Rosa) Pardo, B. (Belén) Sánchez, L. (Laura) Martínez, P. (Paulino) Viñas, A. (Ana) |
author_facet |
Robledo, D. (Diego) Hernández-Urcera, J. (Jorge) Cal, R. (Rosa) Pardo, B. (Belén) Sánchez, L. (Laura) Martínez, P. (Paulino) Viñas, A. (Ana) |
author_sort |
Robledo, D. (Diego) |
title |
Analysis of qPCR reference genes stability determination methods and a practical approach for efficiency calculation on a turbot (Scphthalmus maximus) gonad dataset |
title_short |
Analysis of qPCR reference genes stability determination methods and a practical approach for efficiency calculation on a turbot (Scphthalmus maximus) gonad dataset |
title_full |
Analysis of qPCR reference genes stability determination methods and a practical approach for efficiency calculation on a turbot (Scphthalmus maximus) gonad dataset |
title_fullStr |
Analysis of qPCR reference genes stability determination methods and a practical approach for efficiency calculation on a turbot (Scphthalmus maximus) gonad dataset |
title_full_unstemmed |
Analysis of qPCR reference genes stability determination methods and a practical approach for efficiency calculation on a turbot (Scphthalmus maximus) gonad dataset |
title_sort |
analysis of qpcr reference genes stability determination methods and a practical approach for efficiency calculation on a turbot (scphthalmus maximus) gonad dataset |
publisher |
Centro Oceanográfico de Vigo |
publishDate |
2014 |
url |
http://hdl.handle.net/10508/2045 https://doi.org/10.1186/1471-2164-15-648 |
genre |
Scophthalmus maximus Turbot |
genre_facet |
Scophthalmus maximus Turbot |
op_relation |
http://hdl.handle.net/10508/2045 BMC Genomics, 15. 2014: 648-657 doi:10.1186/1471-2164-15-648 |
op_rights |
Atribución-NoComercial-SinDerivadas 3.0 España http://creativecommons.org/licenses/by-nc-nd/3.0/es/ restrictedAccess |
op_rightsnorm |
CC-BY-NC-ND |
op_doi |
https://doi.org/10.1186/1471-2164-15-648 |
container_title |
BMC Genomics |
container_volume |
15 |
container_issue |
1 |
container_start_page |
648 |
_version_ |
1766188901577261056 |