不同化学类型樟树叶片总RNA提取方法比较  

Comparative research on extraction methods for total RNA from leaves of five chemotypys of Cinnamomum camphora

作　　者：伍艳芳[1] 徐海宁[1] 江香梅[1] 温强[1] 熊振宇[1] 

机构地区：[1]江西省林业科学院,江西省植物生物技术重点实验室,南昌330032

出　　处：《林业科技开发》2013年第4期75-79,共5页China Forestry Science and Technology

基　　金：江西省科技支撑项目(编号:20112BBF60043);江西省科技支撑项目(编号:20113BEA27008)

摘　　要：采用SDS/酸酚法、Trizol试剂法、常规CTAB法及改良的CTAB法,分别提取5种化学类型樟树叶片总RNA,并对提取效果进行了比较分析。结果表明:Trizol法难以提取樟树叶片总RNA,得率很低;SDS/酸酚法和常规CTAB法得到的RNA存在DNA污染,富含蛋白、多糖、多酚等杂质;这3种方法均不适于富含多糖多酚类物质的樟树叶片总RNA的提取。改良的CTAB法能够提取得到樟树5种化学类型叶片总RNA,且在操作过程中,异樟比油樟和脑樟更易提取。此法能有效去除多糖和蛋白,提取得到的RNA 28S和18S rRNA条带清晰,OD260/OD280值为2.0左右,平均产率分别为115.8μg/g FW。经RT-PCR检测,所得总RNA质量高、完整性好、成功率高,可以满足下一步实验的要求,可作为樟树叶片总RNA提取的首选方法。Total RNA from leaves of Cinnamomum camphora were extracted by SDS/phenol extraction, Trizol, common CTAB and the modified methods of CTAB, and the effects of extraction were compared. The results showed that total RNA could not be extracted by Trizol method, the yield was very low and not enough for molecular biological experiment. Total RNA extracted by SDS/phenol and common CTAB extraction ways was immingled by some polysaccharides, protein and DNA. Trizol, SDS/phenol and common CTAB methods were not suitable for total RNA extraction from C. camphora. The improved CTAB method could not only efficiently remove polysaccharides, protein and DNA, but also could extract highquality and high-integrity RNA, as determined by UV-spectroscopic analysis showing the OD260/OD280 at 2.0. The yields of total RNA were 115.8 μg/g FW for the leaf. The results of RT-PCR indicated that this improved CTAB method satisfied the standard for gene cloning and other molecular biological experiments.

关 键 词：樟树 化学类型 RNA提取 CTAB法 RT-PCR 

分 类 号：S791.22[农业科学—林木遗传育种]