NDLI: Identification of suitable endogenous control genes for quantitative RT-PCR analysis of miRNA in bovine solid tissues

Content Provider	Springer Nature Link
Author	Li, Dongwei Liu, Hongyu Li, Yunsheng Yang, Manman Qu, Changqing Zhang, Yunhai Liu, Ya Zhang, Xiaorong
Copyright Year	2014
Abstract	Quantitative reverse transcription polymerase chain reaction (qRT-PCR) has become the preferred technique for studying low-abundance RNA expression. Proper normalization is a critical but often underappreciated aspect of quantitative gene expression analysis; popular endogenous control genes are usually selected with little knowledge of their real suitability. To date, there are very few reports regarding the general validation of endogenous control genes for microRNA (miRNA) expression analysis in bovine tissue. In the present study, eight candidate reference genes (U6, 18S rRNA, GAPDH, ACTB, miR-191, miR-15a, miR-18a, let-7f) were tested for use as normalizers of bovine miRNA in RT-qPCR assays. Their selection was based on publicly available data concerning normalization, hierarchical clustering and sequencing. Three of the genes (miR-191, U6-1 and let-7f) were found to be highly consistent in their expression across eight different bovine solid tissues. It is commonly accepted that gene expression studies should be normalized using more than one endogenous control gene. Based on our results, we propose using the combined results for miR-191, U6-1 and let-7f as the endogenous control for normalization of miRNA levels in qRT-PCR analysis of diverse bovine tissues. This result could act a guideline for future work on bovine miRNA expression.
Starting Page	6475
Ending Page	6480
Page Count	6
File Format	PDF
ISSN	03014851
Journal	Molecular Biology Reports
Volume Number	41
Issue Number	10
e-ISSN	15734978
Language	English
Publisher	Springer Netherlands
Publisher Date	2014-06-29
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Bovine miRNA RT-qPCR Endogenous control Normalization Animal Biochemistry Animal Anatomy / Morphology / Histology
Content Type	Text
Resource Type	Article
Subject	Genetics Medicine Molecular Biology

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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