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An improved method for isolation of RNA from bone
BACKGROUND: Bone physiology is increasingly appreciated as an important contributor to metabolic disorders such as type 2 diabetes. However, progress in understanding the role of bone in determining metabolic health is hampered by the well-described difficulty of obtaining high quality RNA from bone...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3282642/ https://www.ncbi.nlm.nih.gov/pubmed/22260224 http://dx.doi.org/10.1186/1472-6750-12-5 |
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author | Carter, Lauren E Kilroy, Gail Gimble, Jeffrey M Floyd, Z Elizabeth |
author_facet | Carter, Lauren E Kilroy, Gail Gimble, Jeffrey M Floyd, Z Elizabeth |
author_sort | Carter, Lauren E |
collection | PubMed |
description | BACKGROUND: Bone physiology is increasingly appreciated as an important contributor to metabolic disorders such as type 2 diabetes. However, progress in understanding the role of bone in determining metabolic health is hampered by the well-described difficulty of obtaining high quality RNA from bone for gene expression analysis using the currently available approaches. RESULTS: We developed a simple approach to isolate bone RNA that combines pulverizing the bone and the phenol-guanidinium based RNA extraction in a single step while maintaining near-freezing temperatures. This single step method increases the yield of high quality RNA by eight-fold, with RNA integrity numbers ranging from 6.7 to 9.2. CONCLUSIONS: Our streamlined approach substantially increases the yield of high-quality RNA from bone tissue while facilitating safe and efficient processing of multiple samples using readily available platforms. The RNA obtained from this method is suitable for use in gene expression analysis in real-time quantitative PCR, microarray, and next generation sequencing applications. |
format | Online Article Text |
id | pubmed-3282642 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-32826422012-02-21 An improved method for isolation of RNA from bone Carter, Lauren E Kilroy, Gail Gimble, Jeffrey M Floyd, Z Elizabeth BMC Biotechnol Methodology Article BACKGROUND: Bone physiology is increasingly appreciated as an important contributor to metabolic disorders such as type 2 diabetes. However, progress in understanding the role of bone in determining metabolic health is hampered by the well-described difficulty of obtaining high quality RNA from bone for gene expression analysis using the currently available approaches. RESULTS: We developed a simple approach to isolate bone RNA that combines pulverizing the bone and the phenol-guanidinium based RNA extraction in a single step while maintaining near-freezing temperatures. This single step method increases the yield of high quality RNA by eight-fold, with RNA integrity numbers ranging from 6.7 to 9.2. CONCLUSIONS: Our streamlined approach substantially increases the yield of high-quality RNA from bone tissue while facilitating safe and efficient processing of multiple samples using readily available platforms. The RNA obtained from this method is suitable for use in gene expression analysis in real-time quantitative PCR, microarray, and next generation sequencing applications. BioMed Central 2012-01-19 /pmc/articles/PMC3282642/ /pubmed/22260224 http://dx.doi.org/10.1186/1472-6750-12-5 Text en Copyright ©2012 Carter et al; 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 | Methodology Article Carter, Lauren E Kilroy, Gail Gimble, Jeffrey M Floyd, Z Elizabeth An improved method for isolation of RNA from bone |
title | An improved method for isolation of RNA from bone |
title_full | An improved method for isolation of RNA from bone |
title_fullStr | An improved method for isolation of RNA from bone |
title_full_unstemmed | An improved method for isolation of RNA from bone |
title_short | An improved method for isolation of RNA from bone |
title_sort | improved method for isolation of rna from bone |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3282642/ https://www.ncbi.nlm.nih.gov/pubmed/22260224 http://dx.doi.org/10.1186/1472-6750-12-5 |
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