蒙古沙冬青总RNA提取与mRNA分离方法的研究  

Methods for Total RNA Extraction and mRNA isolation from Ammopiptanthus mongolicus

作　　者：林清芳[1] 王存芳[1] 赵欢欢[1] 刘佳杰[1] 王茅雁[1] 

机构地区：[1]内蒙古农业大学生命科学学院,呼和浩特010018

出　　处：《植物遗传资源学报》2011年第3期460-463,共4页Journal of Plant Genetic Resources

基　　金：国家自然科学基金项目(30960158);内蒙古科技计划项目(K32227);国家转基因生物新品种培育重大专项(2009zx08009-019B)

摘　　要：蒙古沙冬青是分布于我国西北荒漠区的常绿旱生阔叶灌木,因其富含多糖和多酚等次生代谢物质,用常规RNA提取方法难以从中获得高质量总RNA。本研究通过在热酚法的RNA提取液中加入高浓度的KAc和β-巯基乙醇,从该植物的不同样品中提取到高质量总RNA,并用筛选到的合适试剂盒分离到高纯度的mRNA。所得到的总RNA和mRNA已被成功应用于基因克隆和SMART全长cDNA文库的构建。Ammopiptanthus mongolicus（Maxim.） is an evergreen broad-leaf shrub in the northwest desert of China.Because of the high content in polysaccharides,polyphenols and other secondary metabolites,it is difficult to extract high-quality total RNA from the plant with conventional methods.In this research,by adding high concentration of potassium acetate and β-mercaptoethanol into the extraction buffer of Hot Phenol Method,we successfully extracted high-quality total RNA from different samples of the plant.We also obtained high-purity mRNA with suitable mRNA isolation kit.Both the total RNA and mRNA have been successfully applied to gene cloning and SMART（Switching Mechanism At 5′end of RNA Transcript） cDNA library construction.

关 键 词：蒙古沙冬青 总RNA提取 mRNA分离 

分 类 号：S793.9[农业科学—林木遗传育种] Q943.2[农业科学—林学]