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Reference gene selection for microRNA expression analysis in the Pacific oyster
Huang, Wen1,2,3,4; Xu, Fei1,3,4; Wang, Wei1,3,4; Li, Li1,2,4; Que, Huayong1,2,3,4; Zhang, Guofan1,2,3,4
2018-09-01
Source PublicationROMANIAN BIOTECHNOLOGICAL LETTERS
ISSN1224-5984
Volume23Issue:5Pages:14043-14050
AbstractReal-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.
KeywordCrassostrea gigas MicroRNA Real-time quantitative PCR Reference gene
DOI10.26327/RBL2018.224
Indexed BySCI
Language英语
Funding ProjectNational Natural Science Foundation of China[31372515] ; Guangxi Natural Science Foundation[2016GXNSFBA380028] ; China Agriculture Research System[CARS-49] ; Taishan Scholars Climbing Program of Shandong
WOS Research AreaBiotechnology & Applied Microbiology
WOS SubjectBiotechnology & Applied Microbiology
WOS IDWOS:000454870500017
PublisherARS DOCENDI
Citation statistics
Document Type期刊论文
Version出版稿
Identifierhttp://ir.qdio.ac.cn/handle/337002/155956
Collection实验海洋生物学重点实验室
Corresponding AuthorQue, Huayong; Zhang, Guofan
Affiliation1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao, Peoples R China
2.Univ Chinese Acad Sci, Beijing, Peoples R China
3.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao, Peoples R China
4.Chinese Acad Sci, Ctr Ocean Mega Sci, Beijing, Peoples R China
First Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences
Corresponding Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences
Recommended Citation
GB/T 7714
Huang, Wen,Xu, Fei,Wang, Wei,et al. Reference gene selection for microRNA expression analysis in the Pacific oyster[J]. ROMANIAN BIOTECHNOLOGICAL LETTERS,2018,23(5):14043-14050.
APA Huang, Wen,Xu, Fei,Wang, Wei,Li, Li,Que, Huayong,&Zhang, Guofan.(2018).Reference gene selection for microRNA expression analysis in the Pacific oyster.ROMANIAN BIOTECHNOLOGICAL LETTERS,23(5),14043-14050.
MLA Huang, Wen,et al."Reference gene selection for microRNA expression analysis in the Pacific oyster".ROMANIAN BIOTECHNOLOGICAL LETTERS 23.5(2018):14043-14050.
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