Reference gene selection for quantitative real-time PCR (qRT-PCR) expression analysis in Galium aparine L.
To accurately evaluate expression levels of target genes, stable internal reference genes is required for normalization of quantitative real-time PCR (qRT-PCR) data. However, there have been no systematical investigation on the stability of reference genes used in the bedstraw weed, Galium aparine L...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2020-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0226668 |
| _version_ | 1819260274705694720 |
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| author | Xu Su Liuyang Lu Yashe Li Congai Zhen Guilei Hu Kun Jiang Yawei Yan Yanbo Xu Geng Wang Mingwang Shi Xiling Chen Baizhong Zhang |
| author_facet | Xu Su Liuyang Lu Yashe Li Congai Zhen Guilei Hu Kun Jiang Yawei Yan Yanbo Xu Geng Wang Mingwang Shi Xiling Chen Baizhong Zhang |
| author_sort | Xu Su |
| collection | DOAJ |
| description | To accurately evaluate expression levels of target genes, stable internal reference genes is required for normalization of quantitative real-time PCR (qRT-PCR) data. However, there have been no systematical investigation on the stability of reference genes used in the bedstraw weed, Galium aparine L. (BGA). In this study, the expression profiles of seven traditionally used reference genes, namely 18S, 28S, ACT, GAPDH, EF1α, RPL7 and TBP in BGA were assessed under both biotic (developmental time and tissue), and abiotic (temperature, regions and herbicide) conditions. Four analytical algorithms (geNorm, Normfinder, BestKeeper and the ΔCt method) were used to analyze the suitability of these genes as internal reference genes. RefFinder, a comprehensive analytical software, was used to rank the overall stability of the candidate genes. The optimal normalization internal control genes were ranked as: 28S and RPL7 were best for all the different experimental conditions (developmental stages, tissues, temperature, regions and herbicide treatment); 28S and RPL7 for developmental stages; TBP and GAPDH for different tissues; 28S and GAPDH were relatively stable for different temperature; 28S and TBP were suitable for herbicide treatment. A specific set of reference genes were recommended for each experimental condition in BGA. |
| first_indexed | 2024-12-23T19:23:18Z |
| format | Article |
| id | doaj.art-e42d88d6b4d74d58b379e5448bbbcdcf |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-23T19:23:18Z |
| publishDate | 2020-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-e42d88d6b4d74d58b379e5448bbbcdcf2022-12-21T17:34:07ZengPublic Library of Science (PLoS)PLoS ONE1932-62032020-01-01152e022666810.1371/journal.pone.0226668Reference gene selection for quantitative real-time PCR (qRT-PCR) expression analysis in Galium aparine L.Xu SuLiuyang LuYashe LiCongai ZhenGuilei HuKun JiangYawei YanYanbo XuGeng WangMingwang ShiXiling ChenBaizhong ZhangTo accurately evaluate expression levels of target genes, stable internal reference genes is required for normalization of quantitative real-time PCR (qRT-PCR) data. However, there have been no systematical investigation on the stability of reference genes used in the bedstraw weed, Galium aparine L. (BGA). In this study, the expression profiles of seven traditionally used reference genes, namely 18S, 28S, ACT, GAPDH, EF1α, RPL7 and TBP in BGA were assessed under both biotic (developmental time and tissue), and abiotic (temperature, regions and herbicide) conditions. Four analytical algorithms (geNorm, Normfinder, BestKeeper and the ΔCt method) were used to analyze the suitability of these genes as internal reference genes. RefFinder, a comprehensive analytical software, was used to rank the overall stability of the candidate genes. The optimal normalization internal control genes were ranked as: 28S and RPL7 were best for all the different experimental conditions (developmental stages, tissues, temperature, regions and herbicide treatment); 28S and RPL7 for developmental stages; TBP and GAPDH for different tissues; 28S and GAPDH were relatively stable for different temperature; 28S and TBP were suitable for herbicide treatment. A specific set of reference genes were recommended for each experimental condition in BGA.https://doi.org/10.1371/journal.pone.0226668 |
| spellingShingle | Xu Su Liuyang Lu Yashe Li Congai Zhen Guilei Hu Kun Jiang Yawei Yan Yanbo Xu Geng Wang Mingwang Shi Xiling Chen Baizhong Zhang Reference gene selection for quantitative real-time PCR (qRT-PCR) expression analysis in Galium aparine L. PLoS ONE |
| title | Reference gene selection for quantitative real-time PCR (qRT-PCR) expression analysis in Galium aparine L. |
| title_full | Reference gene selection for quantitative real-time PCR (qRT-PCR) expression analysis in Galium aparine L. |
| title_fullStr | Reference gene selection for quantitative real-time PCR (qRT-PCR) expression analysis in Galium aparine L. |
| title_full_unstemmed | Reference gene selection for quantitative real-time PCR (qRT-PCR) expression analysis in Galium aparine L. |
| title_short | Reference gene selection for quantitative real-time PCR (qRT-PCR) expression analysis in Galium aparine L. |
| title_sort | reference gene selection for quantitative real time pcr qrt pcr expression analysis in galium aparine l |
| url | https://doi.org/10.1371/journal.pone.0226668 |
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