荒漠甲虫总RNA提取方法的比较与分析  

Comparison and Analysis of the Methods of RNA Isolation for Anatolica Polita Borealis

作　　者：王鹏[1] 热依汗古丽.阿里木 刘忠渊[1] 

机构地区：[1]新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室,新疆乌鲁木齐830046

出　　处：《生物技术》2012年第5期36-39,共4页Biotechnology

基　　金：新疆生物资源基因工程重点实验室开放课题(XJDX0201-2012-01);新疆自治区科技支疆项目(201291304)资助

摘　　要：目的:昆虫总RNA由于其自身结构特点,存在与动物植物完全不同的电泳条带,本文通过对比探索光滑鳖甲总RNA提取的最优方案。方法:实验采用5龄光滑鳖甲幼虫,利用TRIzol、CTAB、热酚法、及BioTeKe Kit、TianGen Kit试剂盒提取方法提取光滑鳖甲总RNA。结果:(1)所采用的实验方法提取的光滑鳖甲总RNA均呈现出与植物、哺乳动物血细胞、哺乳动物组织样总RNA不同的条带,28S弱于18S条带亮度;(2)TRIzol法、CTAB法、改进热酚法所得光滑鳖甲总RNA条带清晰、完整,D260/D280值为1.89～1.98,D260/D230值为1.94～2.09,纯度较好;(3)TRIzol法RNA提取得率为304.3～365.4μg/g,BioTeKe Kit RNA提取得率为73.38～128.22μg/g。结论:综合所有参数,所选取的方法中,TRIzol法可获得高质量、高产量的光滑鳖甲总RNA,可用于RT-PCR、文库构建等高要求RNA的提取。Objective：The total RNA from insects had a large different Electrophoretic bands from animals and plants by its own Structural characteristics. This paper committed to compare and analyze the methods of total RNA for Anatolica Polita borealis ,which we probed the optimal method. Method：Total RNA was isolated from the larvas at the age of 5 of Anatolica Polita borealis with the methods of TRIzol, CTAB, improved heat- phenol and commonly used RNA isolation kits. Result：（ 1 ）The total RNA isolated from Anatolica Polita borealis by the methods we chosed had a special bands hke degraded from plant, the hemecytes and spleen from mammalian. The luminance of band -28S is dimmer than 18S. （2）The total RNA isolated from Anatolica Polita borealis by the method of TRIzol, CTAB and improved heat - phenol was clear and highly - complete. The D260/D280 absorbance ratio of all RNA ranged from 1.89 to 1.98. And their value of D260/D230ranged from 1.94 to 2. 09. It indicated the RNA was highly pure. （3） RNA obtained by the method of TRlznl had highest produc- tivity 304. 3-365.4μg· g-1 ,and BioTeKe Kit method got 73.38 -128. 221μg ·g-l. Condutfion： In all, compared all parameters,the RNA obtained by the method of TRIzol was of the highest quality, highest yields, from the methods checked out in this experiment, and suitable for all downstream applications.

关 键 词：光滑鳖甲 异长足漠甲 幼虫 RNA提取 方法选择与优化 

分 类 号：Q752[生物学—分子生物学]