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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2016
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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. |
format | Online Article Text |
id | pubmed-4928274 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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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