摘要
目的:通过对TRIZOL法加以改进,建立一种快速、有效提取矿化骨组织中总RNA的方法。方法:改进TRIZOL法提取骨组织中的总RNA,在异丙醇沉淀RNA时加入高浓度的盐溶液。通过紫外分光光度法和1%甲醛变性琼脂糖凝胶电泳鉴定所提取的RNA。进而以其逆转录所得的cDNA为模板经PCR扩增OPG基因予以鉴定。结果:改良TRIZOL法获得的总RNA完整,纯度好,产量高。结论:改良TRIZOL法是一种从矿化骨组织中提取总RNA的高效、便捷、可靠的方法。
Objective： To establish a kind of rapid and efficient modified method to isolation total RNA with TRIZOL reagent from mineralized skeletal tissues. Methods： After initial treatment with TRIZOL and chloroform,high concentration saline solution was added in when RNA was precipitated by isopropanol. RNA quality and quantity were determined by spectrophotometer,electrophoresis and RT-PCR. The osteopmtegerin （OPG）gene fragment was amplified through RT-PCR to further test the quality of RNA. Results： Compared with RNA extracted with TRIZOL, RNA extracted using the modified method with no degradation had higher purity and yield. Conclusion： The modified method of RNA isolation with TRIZOL reagent from mineralized skeletal tissues is reliable, efficient and convenient.
出处
《天津医药》
CAS
北大核心
2007年第10期753-754,801,共3页
Tianjin Medical Journal
基金
天津市自然科学基金资助项目(项目编号:043606811)
关键词
RNA
骨和骨组织
逆转录聚合酶链反应
大鼠
RNA bone and bones reverse transcriptase polymerase chain reaction rats
