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Highly efficient method for isolation of total RNA from adipose tissue

BACKGROUND: RNA extraction is a crucial step for monitoring gene expression. Poor RNA quality (including degradation and remaining impurities) can result in misleading results. Isolation of RNA from animal tissues with high lipid content can be challenging. Especially, it is not trivial to isolate h...

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Autor principal: Cirera, Susanna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4225616/
https://www.ncbi.nlm.nih.gov/pubmed/24245791
http://dx.doi.org/10.1186/1756-0500-6-472
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author Cirera, Susanna
author_facet Cirera, Susanna
author_sort Cirera, Susanna
collection PubMed
description BACKGROUND: RNA extraction is a crucial step for monitoring gene expression. Poor RNA quality (including degradation and remaining impurities) can result in misleading results. Isolation of RNA from animal tissues with high lipid content can be challenging. Especially, it is not trivial to isolate high quality RNA with a reasonable yield from adipose tissue. The aim of this study was to provide an optimized protocol for isolating total RNA from adipose tissue. This was achieved by combining the advantages of the two routinely used methods, TRI Reagent® and miRNeasy. FINDINGS: The miRNeasy method results in cleaner samples but more prone to degradation while the TRI Reagent® method results in samples contaminated with salts and solvents but more intact. The new protocol combines the best of both methods resulting in RNA of high quality and suitable for downstream experiments like RT-qPCR, microarrays and high-throughput sequencing. CONCLUSIONS: The current protocol for total RNA isolation from adipose tissue yields sufficient amount of high quality total RNA free of contaminants.
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spelling pubmed-42256162014-11-11 Highly efficient method for isolation of total RNA from adipose tissue Cirera, Susanna BMC Res Notes Technical Note BACKGROUND: RNA extraction is a crucial step for monitoring gene expression. Poor RNA quality (including degradation and remaining impurities) can result in misleading results. Isolation of RNA from animal tissues with high lipid content can be challenging. Especially, it is not trivial to isolate high quality RNA with a reasonable yield from adipose tissue. The aim of this study was to provide an optimized protocol for isolating total RNA from adipose tissue. This was achieved by combining the advantages of the two routinely used methods, TRI Reagent® and miRNeasy. FINDINGS: The miRNeasy method results in cleaner samples but more prone to degradation while the TRI Reagent® method results in samples contaminated with salts and solvents but more intact. The new protocol combines the best of both methods resulting in RNA of high quality and suitable for downstream experiments like RT-qPCR, microarrays and high-throughput sequencing. CONCLUSIONS: The current protocol for total RNA isolation from adipose tissue yields sufficient amount of high quality total RNA free of contaminants. BioMed Central 2013-11-18 /pmc/articles/PMC4225616/ /pubmed/24245791 http://dx.doi.org/10.1186/1756-0500-6-472 Text en Copyright © 2013 Cirera; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 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 cited.
spellingShingle Technical Note
Cirera, Susanna
Highly efficient method for isolation of total RNA from adipose tissue
title Highly efficient method for isolation of total RNA from adipose tissue
title_full Highly efficient method for isolation of total RNA from adipose tissue
title_fullStr Highly efficient method for isolation of total RNA from adipose tissue
title_full_unstemmed Highly efficient method for isolation of total RNA from adipose tissue
title_short Highly efficient method for isolation of total RNA from adipose tissue
title_sort highly efficient method for isolation of total rna from adipose tissue
topic Technical Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4225616/
https://www.ncbi.nlm.nih.gov/pubmed/24245791
http://dx.doi.org/10.1186/1756-0500-6-472
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