Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Carter, Lauren E, Kilroy, Gail, Gimble, Jeffrey M, Floyd, Z Elizabeth
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
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
_version_ 1782224093619159040
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
work_keys_str_mv AT carterlaurene animprovedmethodforisolationofrnafrombone
AT kilroygail animprovedmethodforisolationofrnafrombone
AT gimblejeffreym animprovedmethodforisolationofrnafrombone
AT floydzelizabeth animprovedmethodforisolationofrnafrombone
AT carterlaurene improvedmethodforisolationofrnafrombone
AT kilroygail improvedmethodforisolationofrnafrombone
AT gimblejeffreym improvedmethodforisolationofrnafrombone
AT floydzelizabeth improvedmethodforisolationofrnafrombone