Reference gene selection for microRNA expression analysis in the Pacific oyster
Real-time quantitative polymerase chain reaction (qRT-PCR) is a widely used technique for microRNA (miRNA) expression analysis. The accuracy and reproducibility of this technique depends on the availability of stably expressed reference genes, which is absent in molluscs. Here, based on the microRNA...
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ftchinacasciocas:oai:ir.qdio.ac.cn:337002/155956 2023-05-15T15:57:42+02:00 Reference gene selection for microRNA expression analysis in the Pacific oyster Huang, Wen Xu, Fei Wang, Wei Li, Li Que, Huayong Zhang, Guofan 2018-09-01 http://ir.qdio.ac.cn/handle/337002/155956 http://ir.qdio.ac.cn/handle/337002/155957 https://doi.org/10.26327/RBL2018.224 英语 eng ARS DOCENDI ROMANIAN BIOTECHNOLOGICAL LETTERS http://ir.qdio.ac.cn/handle/337002/155956 http://ir.qdio.ac.cn/handle/337002/155957 doi:10.26327/RBL2018.224 Crassostrea gigas MicroRNA Real-time quantitative PCR Reference gene Biotechnology & Applied Microbiology 期刊论文 2018 ftchinacasciocas https://doi.org/10.26327/RBL2018.224 2022-06-27T05:38:52Z Real-time quantitative polymerase chain reaction (qRT-PCR) is a widely used technique for microRNA (miRNA) expression analysis. The accuracy and reproducibility of this technique depends on the availability of stably expressed reference genes, which is absent in molluscs. Here, based on the microRNA transcriptome data of the Pacific oyster, Crassostrea gigas, candidate reference genes were selected from the stably expressed microRNAs and the commonly used housekeeping genes (U1, U6, and 5.8S). The stability of expression was evaluated and the comprehensive ranking of the reference genes was obtained using two statistical algorithms, geNorm and NormFinder. Specific combinations of reference genes were recommended for the developmental larval samples (DLS) and adult organ samples (AOS), respectively; a combination of cgi-miR-71 and Ul was recommended for DLS and a combination of cgi-let-7, cgi-miR-87, and Ul for AOS, under normal physiological conditions. The effectiveness of the reference gene combinations was further validated by quantitative analysis of two selected miRNAs in the DLS and AOS, respectively. This work was the first systematic analysis of miRNA reference gene selection in molluscs. The reference genes identified in this study will be useful for the functional analysis of miRNA in Crassostrea gigas, and also for the miRNA reference gene selection in other molluscan species. Report Crassostrea gigas Pacific oyster Institute of Oceanology, Chinese Academy of Sciences: IOCAS-IR Pacific |
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Open Polar |
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Institute of Oceanology, Chinese Academy of Sciences: IOCAS-IR |
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ftchinacasciocas |
language |
English |
topic |
Crassostrea gigas MicroRNA Real-time quantitative PCR Reference gene Biotechnology & Applied Microbiology |
spellingShingle |
Crassostrea gigas MicroRNA Real-time quantitative PCR Reference gene Biotechnology & Applied Microbiology Huang, Wen Xu, Fei Wang, Wei Li, Li Que, Huayong Zhang, Guofan Reference gene selection for microRNA expression analysis in the Pacific oyster |
topic_facet |
Crassostrea gigas MicroRNA Real-time quantitative PCR Reference gene Biotechnology & Applied Microbiology |
description |
Real-time quantitative polymerase chain reaction (qRT-PCR) is a widely used technique for microRNA (miRNA) expression analysis. The accuracy and reproducibility of this technique depends on the availability of stably expressed reference genes, which is absent in molluscs. Here, based on the microRNA transcriptome data of the Pacific oyster, Crassostrea gigas, candidate reference genes were selected from the stably expressed microRNAs and the commonly used housekeeping genes (U1, U6, and 5.8S). The stability of expression was evaluated and the comprehensive ranking of the reference genes was obtained using two statistical algorithms, geNorm and NormFinder. Specific combinations of reference genes were recommended for the developmental larval samples (DLS) and adult organ samples (AOS), respectively; a combination of cgi-miR-71 and Ul was recommended for DLS and a combination of cgi-let-7, cgi-miR-87, and Ul for AOS, under normal physiological conditions. The effectiveness of the reference gene combinations was further validated by quantitative analysis of two selected miRNAs in the DLS and AOS, respectively. This work was the first systematic analysis of miRNA reference gene selection in molluscs. The reference genes identified in this study will be useful for the functional analysis of miRNA in Crassostrea gigas, and also for the miRNA reference gene selection in other molluscan species. |
format |
Report |
author |
Huang, Wen Xu, Fei Wang, Wei Li, Li Que, Huayong Zhang, Guofan |
author_facet |
Huang, Wen Xu, Fei Wang, Wei Li, Li Que, Huayong Zhang, Guofan |
author_sort |
Huang, Wen |
title |
Reference gene selection for microRNA expression analysis in the Pacific oyster |
title_short |
Reference gene selection for microRNA expression analysis in the Pacific oyster |
title_full |
Reference gene selection for microRNA expression analysis in the Pacific oyster |
title_fullStr |
Reference gene selection for microRNA expression analysis in the Pacific oyster |
title_full_unstemmed |
Reference gene selection for microRNA expression analysis in the Pacific oyster |
title_sort |
reference gene selection for microrna expression analysis in the pacific oyster |
publisher |
ARS DOCENDI |
publishDate |
2018 |
url |
http://ir.qdio.ac.cn/handle/337002/155956 http://ir.qdio.ac.cn/handle/337002/155957 https://doi.org/10.26327/RBL2018.224 |
geographic |
Pacific |
geographic_facet |
Pacific |
genre |
Crassostrea gigas Pacific oyster |
genre_facet |
Crassostrea gigas Pacific oyster |
op_relation |
ROMANIAN BIOTECHNOLOGICAL LETTERS http://ir.qdio.ac.cn/handle/337002/155956 http://ir.qdio.ac.cn/handle/337002/155957 doi:10.26327/RBL2018.224 |
op_doi |
https://doi.org/10.26327/RBL2018.224 |
_version_ |
1766393387464785920 |