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Open Access Highly Accessed Methodology article

An improved method for isolation of RNA from bone

Lauren E Carter1, Gail Kilroy1, Jeffrey M Gimble2 and Z Elizabeth Floyd1*

Author Affiliations

1 Ubiquitin Biology Laboratory, Pennington Biomedical Research Center, 6400 Perkins Road, Baton Rouge, LA 70808 USA

2 Stem Cell Biology Laboratory, Pennington Biomedical Research Center, 6400 Perkins Road, Baton Rouge, LA 70808 USA

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BMC Biotechnology 2012, 12:5  doi:10.1186/1472-6750-12-5

Published: 19 January 2012

Abstract

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.