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Variation of Expression Levels of Seven Housekeeping Genes at Different Life-History Stages in Porphyra yezoensis

In order to identify the optimal internal control for relative real-time PCR when studying target gene expression in the red alga Porphyra yezoensis, we quantified the expression of seven housekeeping genes (18S ribosomal RNA, 30S ribosomal protein S8, Polyubiquitin-2, Glyceraldehyde-3-phosphate deh...

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Autores principales: Wu, Xiaojie, Huang, Aiyou, Xu, Meiling, Wang, Chao, Jia, Zhaojun, Wang, Guangce, Niu, Jianfeng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3630157/
https://www.ncbi.nlm.nih.gov/pubmed/23637763
http://dx.doi.org/10.1371/journal.pone.0060740
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author Wu, Xiaojie
Huang, Aiyou
Xu, Meiling
Wang, Chao
Jia, Zhaojun
Wang, Guangce
Niu, Jianfeng
author_facet Wu, Xiaojie
Huang, Aiyou
Xu, Meiling
Wang, Chao
Jia, Zhaojun
Wang, Guangce
Niu, Jianfeng
author_sort Wu, Xiaojie
collection PubMed
description In order to identify the optimal internal control for relative real-time PCR when studying target gene expression in the red alga Porphyra yezoensis, we quantified the expression of seven housekeeping genes (18S ribosomal RNA, 30S ribosomal protein S8, Polyubiquitin-2, Glyceraldehyde-3-phosphate dehydrogenase, Elongation factor 1-alpha, Beta-tubulin and Actin 3) at different life-history stages. Absolute quantification was done by normalization to total RNA quantity and by normalization to genomic DNA quantity. We used these two normalization approaches, comparing the differences of expression levels of all candidate housekeeping genes between any two generations and across three life-history stages (filamentous sporophytes, leafy gametophytes and conchospores). We found GAPDH had the best stability in all cases and we recommend that GAPDH be considered as a potential internal control for gene expression studies at different life-history stages in P. yezoensis.
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spelling pubmed-36301572013-05-01 Variation of Expression Levels of Seven Housekeeping Genes at Different Life-History Stages in Porphyra yezoensis Wu, Xiaojie Huang, Aiyou Xu, Meiling Wang, Chao Jia, Zhaojun Wang, Guangce Niu, Jianfeng PLoS One Research Article In order to identify the optimal internal control for relative real-time PCR when studying target gene expression in the red alga Porphyra yezoensis, we quantified the expression of seven housekeeping genes (18S ribosomal RNA, 30S ribosomal protein S8, Polyubiquitin-2, Glyceraldehyde-3-phosphate dehydrogenase, Elongation factor 1-alpha, Beta-tubulin and Actin 3) at different life-history stages. Absolute quantification was done by normalization to total RNA quantity and by normalization to genomic DNA quantity. We used these two normalization approaches, comparing the differences of expression levels of all candidate housekeeping genes between any two generations and across three life-history stages (filamentous sporophytes, leafy gametophytes and conchospores). We found GAPDH had the best stability in all cases and we recommend that GAPDH be considered as a potential internal control for gene expression studies at different life-history stages in P. yezoensis. Public Library of Science 2013-04-18 /pmc/articles/PMC3630157/ /pubmed/23637763 http://dx.doi.org/10.1371/journal.pone.0060740 Text en © 2013 Wu et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Wu, Xiaojie
Huang, Aiyou
Xu, Meiling
Wang, Chao
Jia, Zhaojun
Wang, Guangce
Niu, Jianfeng
Variation of Expression Levels of Seven Housekeeping Genes at Different Life-History Stages in Porphyra yezoensis
title Variation of Expression Levels of Seven Housekeeping Genes at Different Life-History Stages in Porphyra yezoensis
title_full Variation of Expression Levels of Seven Housekeeping Genes at Different Life-History Stages in Porphyra yezoensis
title_fullStr Variation of Expression Levels of Seven Housekeeping Genes at Different Life-History Stages in Porphyra yezoensis
title_full_unstemmed Variation of Expression Levels of Seven Housekeeping Genes at Different Life-History Stages in Porphyra yezoensis
title_short Variation of Expression Levels of Seven Housekeeping Genes at Different Life-History Stages in Porphyra yezoensis
title_sort variation of expression levels of seven housekeeping genes at different life-history stages in porphyra yezoensis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3630157/
https://www.ncbi.nlm.nih.gov/pubmed/23637763
http://dx.doi.org/10.1371/journal.pone.0060740
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