Defining quantification methods and optimizing protocols for microarray hybridization of circulating microRNAs

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• dc.contributor.author Garcia-Elias Heras, Annaca
• dc.contributor.author Alloza, Leonorca
• dc.contributor.author Puigdecanet Riubugent, Eulàliaca
• dc.contributor.author Nonell Mazelon, Lara, 1972-ca
• dc.contributor.author Tajes Orduña, Martaca
• dc.contributor.author Curado, Joaoca
• dc.contributor.author Enjuanes Grau, Cristinaca
• dc.contributor.author Díaz, Oscarca
• dc.contributor.author Bruguera-Cortada, Jordica
• dc.contributor.author Martí-Almor, Julioca
• dc.contributor.author Comín Colet, Josepca
• dc.contributor.author Benito Villabriga, Begoñaca
• dc.date.accessioned 2018-07-25T06:44:37Z
• dc.date.available 2018-07-25T06:44:37Z
• dc.date.issued 2017
• dc.description.abstract MicroRNAs (miRNAs) have emerged as promising biomarkers of disease. Their potential use in clinical practice requires standardized protocols with very low miRNA concentrations, particularly in plasma samples. Here we tested the most appropriate method for miRNA quantification and validated the performance of a hybridization platform using lower amounts of starting RNA. miRNAs isolated from human plasma and from a reference sample were quantified using four platforms and profiled with hybridization arrays and RNA sequencing (RNA-seq). Our results indicate that the Infinite® 200 PRO Nanoquant and Nanodrop 2000 spectrophotometers magnified the miRNA concentration by detecting contaminants, proteins, and other forms of RNA. The Agilent 2100 Bioanalyzer PicoChip and SmallChip gave valuable information on RNA profile but were not a reliable quantification method for plasma samples. The Qubit® 2.0 Fluorometer provided the most accurate quantification of miRNA content, although RNA-seq confirmed that only ~58% of small RNAs in plasma are true miRNAs. On the other hand, reducing the starting RNA to 70% of the recommended amount for miRNA profiling with arrays yielded results comparable to those obtained with the full amount, whereas a 50% reduction did not. These findings provide important clues for miRNA determination in human plasma samples.
• dc.format.mimetype application/pdf
• dc.identifier.citation Garcia-Elias A, Alloza L, Puigdecanet E, Nonell L, Tajes M, Curado J et al. Defining quantification methods and optimizing protocols for microarray hybridization of circulating microRNAs. Sci Rep. 2017 Aug 10;7(1):7725. DOI: 10.1038/s41598-017-08134-3
• dc.identifier.doi http://dx.doi.org/10.1038/s41598-017-08134-3
• dc.identifier.issn 2045-2322
• dc.identifier.uri http://hdl.handle.net/10230/35257
• dc.language.iso eng
• dc.publisher Nature Publishing Groupca
• dc.relation.ispartof Scientific Reports. 2017 Aug 10;7(1):7725
• dc.rights Copyright © The Author(s) 2017. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
• dc.rights.accessRights info:eu-repo/semantics/openAccess
• dc.rights.uri http://creativecommons.org/licenses/by/4.0/
• dc.subject.other Marcadors bioquímics
• dc.title Defining quantification methods and optimizing protocols for microarray hybridization of circulating microRNAsca
• dc.type info:eu-repo/semantics/article
• dc.type.version info:eu-repo/semantics/publishedVersion