Abstract:

BACKGROUND: There are no ideal methods to extract total RNA from bone tissues.
OBJECTIVE: To explore an efficient and quick method of acquiring total RNA from bone tissue.
METHODS: Ten healthy rabbits were divided into experimental group and control group on average. The femoral heads of experimental group were removed by rongeur which has been disinfected, then stored in liquid nitrogen after quick-freeze. Normal way was used to get control group’s femoral heads. Experimental group’s femoral head was immediately placed in mortar that has been precooled by liquid nitrogen. The bone was grinded iteratively in mortar until it became bone powder, the powder was transferred into a homogenizer which has been precooled, added Trizol, centrifuged at 4 ℃ after fully homogenized to get supernatant. Then chloroform and other organic solvents were added, centrifuged, and then the total RNA was separated from DNA, protein and other tissues. Control group’s RNA was extracted by traditional Trizols method. The concentration, purity and productive rate were measured by ultraviolet spectrophotometer. Finally, denaturing agarose-formalhyde gel electrophoresis was used to observe if the two bands (28 S,18 S) of RNA were clear, RNA was degradated and with/without DNA contamination.
RESULTS AND CONCLUSION: The extracted RNA were in high purity without DNA and protein contamination; The result of degenerated formaldehyde electrophoresis shows that the 28 S and 18 S bands were clear and the ratio of them was about 1:1, which confirmed that RNA was complete with no degradation. The findings from the present study show that this method is a rapid and efficient purification method for gaining total RNA form bone tissue, and it can be used for analyzing molecular biology of bone tissue.