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Two-tailed RT-qPCR panel for quality control of circulating microRNA studies
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SYSNO ASEP 0505062 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Two-tailed RT-qPCR panel for quality control of circulating microRNA studies Author(s) Androvič, Peter (BTO-N)
Romanyuk, Nataliya (UEM-P) RID
Machová-Urdzíková, Lucia (UEM-P) ORCID
Rohlová, Eva (BTO-N)
Kubista, Mikael (BTO-N) RID
Valihrach, Lukáš (BTO-N) RID, ORCIDNumber of authors 6 Article number 4255 Source Title Scientific Reports. - : Nature Publishing Group - ISSN 2045-2322
Roč. 9, MAR 12 2019 (2019)Number of pages 9 s. Language eng - English Country GB - United Kingdom Keywords PLASMA ; RNA ; STANDARDIZATION ; BIOMARKERS Subject RIV EB - Genetics ; Molecular Biology OECD category Biochemistry and molecular biology Subject RIV - cooperation Institute of Experimental Medicine - Genetics ; Molecular Biology R&D Projects GA18-21942S GA ČR - Czech Science Foundation (CSF) ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Open access Institutional support BTO-N - RVO:86652036 ; UEM-P - RVO:68378041 UT WOS 000460922200091 EID SCOPUS 85062845745 DOI 10.1038/s41598-019-40513-w Annotation Circulating cell-free microRNAs are promising candidates for minimally invasive clinical biomarkers for the diagnosis, prognosis and monitoring of many human diseases. Despite substantial efforts invested in the field, the research so far has failed to deliver expected results. One of the contributing factors is general lack of agreement between various studies, partly due to the considerable technical challenges accompanying the workflow. Pre-analytical variables including sample collection, RNA isolation, and quantification are sources of bias that may hamper biological interpretation of the results. Here, we present a Two-tailed RT-qPCR panel for quality control, monitoring of technical performance, and optimization of microRNA profiling experiments from biofluid samples. The Two-tailed QC (quality control) panel is based on two sets of synthetic spike-in molecules and three endogenous microRNAs that are quantified with the highly specific Two-tailed RT-qPCR technology. The QC panel is a cost-effective way to assess quality of isolated microRNA, degree of inhibition, and erythrocyte contamination to ensure technical soundness of the obtained results. We provide assay sequences, detailed experimental protocol and guide to data interpretation. The application of the QC panel is demonstrated on the optimization of RNA isolation from biofluids with the miRNeasy Serum/Plasma Advanced Kit (Qiagen). Workplace Institute of Biotechnology Contact Monika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700 Year of Publishing 2020 Electronic address https://www.nature.com/articles/s41598-019-40513-w
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