红树植物叶片总RNA提取方法比较与优化  

Analysis and improvement of high-quality RNA extraction in leaves of mangrove plants

作　　者：宋晖[1,2] 王友绍[1,3] 

机构地区：[1]中国科学院南海海洋研究所热带海洋环境动力学重点实验室,广州510301 [2]中国科学院研究生院,北京100039 [3]中国科学院大亚湾海洋生物综合试验站,深圳518121

出　　处：《生态科学》2011年第2期201-206,共6页Ecological Science

基　　金："十一五"国家科技支撑计划重点项(2009BADB2B0606);国家自然科学基金项目(41076070);中国科学院知识创新工程重要方向性项目(KZCX2-YW-Q07-02;KSCX2-EW-G-12C;KSCX2-SW-132)

摘　　要：分别采用三种植物RNA提取试剂盒、改良的CTAB-LiCl法和CTAB-异丙醇法五种方法提取四种红树植物叶片总RNA,并用紫外光谱、琼脂糖凝胶电泳、cDNA合成和real-time PCR等手段对提取效果进行鉴定。反复试验表明:五种方法对白骨壤Aricenniamarina叶片总RNA提取均有良好的效果。TiangenRNA提取试剂能够成功提取秋茄Kandeliacandel、桐花树Aegicerascorniculatum、白骨壤的总RNA,提取的木榄Bruguieragymnorrhiza总RNA浓度低、杂质较多,且稳定性较差,在对该方法改良后则能够成功提取木榄的总RNA。Invitrogen试剂盒对木榄和秋茄叶片均有良好的提取效果,但是对桐花树提取的RNA易受到蛋白的污染。CTAB-LiCl法和CTAB-异丙醇法提取的RNA总产量偏低,提取时间较长,且实时定量结果表明,两种方法反转录效率较试剂盒方法也偏低。Five extraction methods including Takara RNA, Tiangen kit, Invitrogen kit, modified CTAB-LiC1 and CTAB- isopropyl alcohol method were compared for extracting total RNA from the leaves of four mangrove plants （Bruguiera gymnorrhiza, Kandelia candel, Aegiceras corniculatum, and Aricennia marina）. The five methods were all suitable for total RNA extraction in leaves of A. marina. Tiangen kit was successfully used to extract the RNA in leaves ofA.corniculatum, A.marina, and K. candel, but failed to extract the RNA in leaves ofB. gymnorrihiza. Invitrogen kit extracted high quality RNA ofB. gymnorrihiza and K.candel, but could not remove the protein of leaves ofA. corniculatum. The RNA amount extracted using CTAB-LiC1 and improved CTAB-isopropyl alcohol methods was lower than using RNA extraction kits; besides, the efficiency of reverse transcription PCR of CTAB-LiC1 and CTAB- isopropyl alcohol methods was lower.

关 键 词：红树植物 总RNA 提取 方法比较 

分 类 号：Q95-3[生物学—动物学]