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Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells
BACKGROUND: microRNAs (miRNAs) play crucial roles in major biological processes and their deregulations are often associated with human malignancies. As such, they represent appealing candidates as targets of innovative therapies. Another interesting aspect of their biology is that they are present...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4041998/ https://www.ncbi.nlm.nih.gov/pubmed/24885883 http://dx.doi.org/10.1186/1471-2164-15-395 |
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author | Monleau, Marjorie Bonnel, Sophie Gostan, Thierry Blanchard, Dominique Courgnaud, Valérie Lecellier, Charles-Henri |
author_facet | Monleau, Marjorie Bonnel, Sophie Gostan, Thierry Blanchard, Dominique Courgnaud, Valérie Lecellier, Charles-Henri |
author_sort | Monleau, Marjorie |
collection | PubMed |
description | BACKGROUND: microRNAs (miRNAs) play crucial roles in major biological processes and their deregulations are often associated with human malignancies. As such, they represent appealing candidates as targets of innovative therapies. Another interesting aspect of their biology is that they are present in various biological fluids where, advantageously, they appear to be very stable. A plethora of studies have now reported their potential as biomarkers that can be used in diagnosis, prognosis and/or theranostic issues. However, the application of circulating miRNAs in clinical practices still requires the identification of highly efficient, robust and reproducible methods for their isolation from biological samples. In that context, we performed an independent cross-comparison of three commercially available RNA extraction kits for miRNAs isolation from human blood samples (Qiagen and Norgen kits as well as the new NucleoSpin miRNAs Plasma kit from Macherey-Nagel). miRNAs were further profiled using the Taqman Low Density Array technology. RESULTS: We found that, although these 3 kits had equal performances in extracting miRNAs from peripheral blood mononuclear cells, the Macherey-Nagel kit presented several advantages when isolating miRNAs from sera. Besides, our results have indicated that, depending on the quantity of the biological samples used, the extraction procedure directly impacted on the G/C composition of the miRNAs detected. CONCLUSION: Overall, our study contributes to the definition of a reliable framework for profiling circulating miRNAs. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1471-2164-15-395) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4041998 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-40419982014-06-06 Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells Monleau, Marjorie Bonnel, Sophie Gostan, Thierry Blanchard, Dominique Courgnaud, Valérie Lecellier, Charles-Henri BMC Genomics Methodology Article BACKGROUND: microRNAs (miRNAs) play crucial roles in major biological processes and their deregulations are often associated with human malignancies. As such, they represent appealing candidates as targets of innovative therapies. Another interesting aspect of their biology is that they are present in various biological fluids where, advantageously, they appear to be very stable. A plethora of studies have now reported their potential as biomarkers that can be used in diagnosis, prognosis and/or theranostic issues. However, the application of circulating miRNAs in clinical practices still requires the identification of highly efficient, robust and reproducible methods for their isolation from biological samples. In that context, we performed an independent cross-comparison of three commercially available RNA extraction kits for miRNAs isolation from human blood samples (Qiagen and Norgen kits as well as the new NucleoSpin miRNAs Plasma kit from Macherey-Nagel). miRNAs were further profiled using the Taqman Low Density Array technology. RESULTS: We found that, although these 3 kits had equal performances in extracting miRNAs from peripheral blood mononuclear cells, the Macherey-Nagel kit presented several advantages when isolating miRNAs from sera. Besides, our results have indicated that, depending on the quantity of the biological samples used, the extraction procedure directly impacted on the G/C composition of the miRNAs detected. CONCLUSION: Overall, our study contributes to the definition of a reliable framework for profiling circulating miRNAs. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1471-2164-15-395) contains supplementary material, which is available to authorized users. BioMed Central 2014-05-23 /pmc/articles/PMC4041998/ /pubmed/24885883 http://dx.doi.org/10.1186/1471-2164-15-395 Text en © Monleau et al.; licensee BioMed Central Ltd. 2014 This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Methodology Article Monleau, Marjorie Bonnel, Sophie Gostan, Thierry Blanchard, Dominique Courgnaud, Valérie Lecellier, Charles-Henri Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells |
title | Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells |
title_full | Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells |
title_fullStr | Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells |
title_full_unstemmed | Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells |
title_short | Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells |
title_sort | comparison of different extraction techniques to profile micrornas from human sera and peripheral blood mononuclear cells |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4041998/ https://www.ncbi.nlm.nih.gov/pubmed/24885883 http://dx.doi.org/10.1186/1471-2164-15-395 |
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