摘要
为了改进经典Trizol法,我们建立了一种有效提取矿化骨组织中总RNA的新方法。在Trizol裂解时加入等体积水饱和酚,在异丙醇沉淀时加入高浓度的盐溶液,并重复氯仿抽提1次。通过紫外分光光度法和1%甲醛变性凝胶电泳法分析提取RNA,以其逆转录cDNA为模板,经PCR扩增雌激素受体(ER)基因予以鉴定。结果,本方法获得的总RNA A260/A280达到1.85以上,电泳条带显示RNA完整,RT-PCR结果显示其纯度较好。结果表明,改良Trizol法是一种从矿化骨组织中提取总RNA的高效便捷的方法。
To establish as efficient modified method for isolation of total RNA with Trizol regent from the bone tissues.Equal volume of both satuated water phenol was added when initial treatment with Trizol,After two times of treatment with chloroform,high concentration saline solution was added in when RNA was precipitated by isopropanol.The quality and quantity of RNA were determined by ultraviolet spectrophotometer,electrophoresis and RT-PCR.The estrogen receptor（ER） gene fragment was amplified through RT-PCR to further test the quality of RNA.Satuated RNA extrated by the modified method with no degradation had higher purity and yield.The modified method of RNA isolation from bone tissue is efficient and convenient.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2010年第10期1368-1370,共3页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(30671546)
