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Optimized methods for extracting circulating small RNAs from long-term stored equine samples

Circulating miRNAs in body fluids, particularly serum, are promising candidates for future routine biomarker profiling in various pathologic conditions in human and veterinary medicine. However, reliable standardized methods for miRNA extraction from equine serum and fresh or archived whole blood ar...

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Detalles Bibliográficos
Autores principales: Unger, Lucia, Fouché, Nathalie, Leeb, Tosso, Gerber, Vincent, Pacholewska, Alicja
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4928274/
https://www.ncbi.nlm.nih.gov/pubmed/27356979
http://dx.doi.org/10.1186/s13028-016-0224-5
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author Unger, Lucia
Fouché, Nathalie
Leeb, Tosso
Gerber, Vincent
Pacholewska, Alicja
author_facet Unger, Lucia
Fouché, Nathalie
Leeb, Tosso
Gerber, Vincent
Pacholewska, Alicja
author_sort Unger, Lucia
collection PubMed
description Circulating miRNAs in body fluids, particularly serum, are promising candidates for future routine biomarker profiling in various pathologic conditions in human and veterinary medicine. However, reliable standardized methods for miRNA extraction from equine serum and fresh or archived whole blood are sorely lacking. We systematically compared various miRNA extraction methods from serum and whole blood after short and long-term storage without addition of RNA stabilizing additives prior to freezing. Time of storage at room temperature prior to freezing did not affect miRNA quality in serum. Furthermore, we showed that miRNA of NGS-sufficient quality can be recovered from blood samples after >10 years of storage at −80 °C. This allows retrospective analyses of miRNAs from archived samples. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13028-016-0224-5) contains supplementary material, which is available to authorized users.
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spelling pubmed-49282742016-06-30 Optimized methods for extracting circulating small RNAs from long-term stored equine samples Unger, Lucia Fouché, Nathalie Leeb, Tosso Gerber, Vincent Pacholewska, Alicja Acta Vet Scand Brief Communication Circulating miRNAs in body fluids, particularly serum, are promising candidates for future routine biomarker profiling in various pathologic conditions in human and veterinary medicine. However, reliable standardized methods for miRNA extraction from equine serum and fresh or archived whole blood are sorely lacking. We systematically compared various miRNA extraction methods from serum and whole blood after short and long-term storage without addition of RNA stabilizing additives prior to freezing. Time of storage at room temperature prior to freezing did not affect miRNA quality in serum. Furthermore, we showed that miRNA of NGS-sufficient quality can be recovered from blood samples after >10 years of storage at −80 °C. This allows retrospective analyses of miRNAs from archived samples. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13028-016-0224-5) contains supplementary material, which is available to authorized users. BioMed Central 2016-06-29 /pmc/articles/PMC4928274/ /pubmed/27356979 http://dx.doi.org/10.1186/s13028-016-0224-5 Text en © Unger et al 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 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 Brief Communication
Unger, Lucia
Fouché, Nathalie
Leeb, Tosso
Gerber, Vincent
Pacholewska, Alicja
Optimized methods for extracting circulating small RNAs from long-term stored equine samples
title Optimized methods for extracting circulating small RNAs from long-term stored equine samples
title_full Optimized methods for extracting circulating small RNAs from long-term stored equine samples
title_fullStr Optimized methods for extracting circulating small RNAs from long-term stored equine samples
title_full_unstemmed Optimized methods for extracting circulating small RNAs from long-term stored equine samples
title_short Optimized methods for extracting circulating small RNAs from long-term stored equine samples
title_sort optimized methods for extracting circulating small rnas from long-term stored equine samples
topic Brief Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4928274/
https://www.ncbi.nlm.nih.gov/pubmed/27356979
http://dx.doi.org/10.1186/s13028-016-0224-5
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