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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Centro Oceanográfico de Vigo
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ftcsic:oai:digital.csic.es:10261/315449 2024-02-11T10:08:26+01:00 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) 2014-08-22T09:01:03Z http://hdl.handle.net/10508/2045 http://hdl.handle.net/10261/315449 https://doi.org/10.1186/1471-2164-15-648 en eng Centro Oceanográfico de Vigo http://hdl.handle.net/10508/2045 http://hdl.handle.net/10261/315449 doi:10.1186/1471-2164-15-648 BMC Genomics, 15. 2014: 648-657 20012 open qPCR Reference genes Amplification efficiency Turbot Scophthalmus maximus research article VoR SI 2014 ftcsic https://doi.org/10.1186/1471-2164-15-648 2024-01-16T11:44:17Z 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 Article in Journal/Newspaper Scophthalmus maximus Turbot Digital.CSIC (Spanish National Research Council) BMC Genomics 15 1 |
institution |
Open Polar |
collection |
Digital.CSIC (Spanish National Research Council) |
op_collection_id |
ftcsic |
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 |
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 http://hdl.handle.net/10261/315449 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 http://hdl.handle.net/10261/315449 doi:10.1186/1471-2164-15-648 BMC Genomics, 15. 2014: 648-657 20012 |
op_rights |
open |
op_doi |
https://doi.org/10.1186/1471-2164-15-648 |
container_title |
BMC Genomics |
container_volume |
15 |
container_issue |
1 |
_version_ |
1790607754647306240 |