Reference gene validation for quantification of gene expression during ovarian development of turbot (Scophthalmus maximus)
Abstract Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) is a powerful and sensitive method used in gene expression analysis. Suitable reference genes, which are stable under all experimental circumstances and tissues significantly improve the accuracy of qRT-PCR dat...
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Online Access: | http://dx.doi.org/10.1038/s41598-020-57633-3 http://www.nature.com/articles/s41598-020-57633-3.pdf http://www.nature.com/articles/s41598-020-57633-3 |
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crspringernat:10.1038/s41598-020-57633-3 2023-05-15T18:15:51+02:00 Reference gene validation for quantification of gene expression during ovarian development of turbot (Scophthalmus maximus) Gao, Yunhong Gao, Yuntao Huang, Bin Meng, Zhen Jia, Yudong National Natural Science Foundation of China Earmarked Fund for China Agriculture Research System 2020 http://dx.doi.org/10.1038/s41598-020-57633-3 http://www.nature.com/articles/s41598-020-57633-3.pdf http://www.nature.com/articles/s41598-020-57633-3 en eng Springer Science and Business Media LLC https://creativecommons.org/licenses/by/4.0 https://creativecommons.org/licenses/by/4.0 CC-BY Scientific Reports volume 10, issue 1 ISSN 2045-2322 Multidisciplinary journal-article 2020 crspringernat https://doi.org/10.1038/s41598-020-57633-3 2022-01-04T16:15:28Z Abstract Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) is a powerful and sensitive method used in gene expression analysis. Suitable reference genes, which are stable under all experimental circumstances and tissues significantly improve the accuracy of qRT-PCR data. In this study, the stability of six genes, namely, 18S ribosomal RNA (18 s ), beta-actin ( actb ), elongation factor 1-alpha ( ef1α ), glyceraldehyde-3-phosphate-dehydrogenase ( gapdh ), cathepsin D ( ctsd ), and beta-2-microglobulin ( b2m ) were evaluated as potential references for qRT-PCR analysis. The genes were examined in the hypothalamus-pituitary-ovary-liver (HPOL) axis throughout turbot ovarian development via using the geNorm, NormFinder and BestKeeper algorithms. Results showed that the most stable reference genes were ef1α , actb , and ctsd in the hypothalamus, pituitary, ovary and liver, respectively. The best-suited gene combinations for normalization were 18 s , ef1α , and ctsd in the hypothalamus; actb , ctsd , and 18 s in the pituitary; actb , and ctsd in the ovary; gapdh and ctsd in the liver. Moreover, the expression profile of estrogen receptor α ( erα ) manifested no significant difference normalization to the aforementioned best-suited gene during turbot ovarian development. However, no single gene or pair of genes is suitable as an internal control and account for the amplification differences among the four tissues during ovarian development. In summary, these results provide a basic data for the optimal reference gene selection and obtain highly accurate normalization of qRT-PCR data in HPOL axis-related gene expression analysis during turbot ovarian development. Article in Journal/Newspaper Scophthalmus maximus Turbot Springer Nature (via Crossref) Scientific Reports 10 1 |
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Multidisciplinary |
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Multidisciplinary Gao, Yunhong Gao, Yuntao Huang, Bin Meng, Zhen Jia, Yudong Reference gene validation for quantification of gene expression during ovarian development of turbot (Scophthalmus maximus) |
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Multidisciplinary |
description |
Abstract Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) is a powerful and sensitive method used in gene expression analysis. Suitable reference genes, which are stable under all experimental circumstances and tissues significantly improve the accuracy of qRT-PCR data. In this study, the stability of six genes, namely, 18S ribosomal RNA (18 s ), beta-actin ( actb ), elongation factor 1-alpha ( ef1α ), glyceraldehyde-3-phosphate-dehydrogenase ( gapdh ), cathepsin D ( ctsd ), and beta-2-microglobulin ( b2m ) were evaluated as potential references for qRT-PCR analysis. The genes were examined in the hypothalamus-pituitary-ovary-liver (HPOL) axis throughout turbot ovarian development via using the geNorm, NormFinder and BestKeeper algorithms. Results showed that the most stable reference genes were ef1α , actb , and ctsd in the hypothalamus, pituitary, ovary and liver, respectively. The best-suited gene combinations for normalization were 18 s , ef1α , and ctsd in the hypothalamus; actb , ctsd , and 18 s in the pituitary; actb , and ctsd in the ovary; gapdh and ctsd in the liver. Moreover, the expression profile of estrogen receptor α ( erα ) manifested no significant difference normalization to the aforementioned best-suited gene during turbot ovarian development. However, no single gene or pair of genes is suitable as an internal control and account for the amplification differences among the four tissues during ovarian development. In summary, these results provide a basic data for the optimal reference gene selection and obtain highly accurate normalization of qRT-PCR data in HPOL axis-related gene expression analysis during turbot ovarian development. |
author2 |
National Natural Science Foundation of China Earmarked Fund for China Agriculture Research System |
format |
Article in Journal/Newspaper |
author |
Gao, Yunhong Gao, Yuntao Huang, Bin Meng, Zhen Jia, Yudong |
author_facet |
Gao, Yunhong Gao, Yuntao Huang, Bin Meng, Zhen Jia, Yudong |
author_sort |
Gao, Yunhong |
title |
Reference gene validation for quantification of gene expression during ovarian development of turbot (Scophthalmus maximus) |
title_short |
Reference gene validation for quantification of gene expression during ovarian development of turbot (Scophthalmus maximus) |
title_full |
Reference gene validation for quantification of gene expression during ovarian development of turbot (Scophthalmus maximus) |
title_fullStr |
Reference gene validation for quantification of gene expression during ovarian development of turbot (Scophthalmus maximus) |
title_full_unstemmed |
Reference gene validation for quantification of gene expression during ovarian development of turbot (Scophthalmus maximus) |
title_sort |
reference gene validation for quantification of gene expression during ovarian development of turbot (scophthalmus maximus) |
publisher |
Springer Science and Business Media LLC |
publishDate |
2020 |
url |
http://dx.doi.org/10.1038/s41598-020-57633-3 http://www.nature.com/articles/s41598-020-57633-3.pdf http://www.nature.com/articles/s41598-020-57633-3 |
genre |
Scophthalmus maximus Turbot |
genre_facet |
Scophthalmus maximus Turbot |
op_source |
Scientific Reports volume 10, issue 1 ISSN 2045-2322 |
op_rights |
https://creativecommons.org/licenses/by/4.0 https://creativecommons.org/licenses/by/4.0 |
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CC-BY |
op_doi |
https://doi.org/10.1038/s41598-020-57633-3 |
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Scientific Reports |
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