Background: Bone physiology is increasingly appreciated as an important contributor to metabolic disorders such\r\nas type 2 diabetes. However, progress in understanding the role of bone in determining metabolic health is\r\nhampered by the well-described difficulty of obtaining high quality RNA from bone for gene expression analysis\r\nusing the currently available approaches.\r\nResults: We developed a simple approach to isolate bone RNA that combines pulverizing the bone and the\r\nphenol-guanidinium based RNA extraction in a single step while maintaining near-freezing temperatures. This\r\nsingle step method increases the yield of high quality RNA by eight-fold, with RNA integrity numbers ranging from\r\n6.7 to 9.2.\r\nConclusions: Our streamlined approach substantially increases the yield of high-quality RNA from bone tissue\r\nwhile facilitating safe and efficient processing of multiple samples using readily available platforms. The RNA\r\nobtained from this method is suitable for use in gene expression analysis in real-time quantitative PCR, microarray,\r\nand next generation sequencing applications
Loading....