土壤RNA提取方法研究进展  

Progress of Researches on Extraction Methods of Soil RNA

作　　者：刘华[1] 申天琳[2,3] 李学坤[2] 付合才[2,3] 戴九兰[1] 

机构地区：[1]山东大学环境研究院,济南250100 [2]山东大学生命科学学院生态学与生物多样性研究所,济南250100 [3]山东大学山东省植被生态工程技术中心,济南250100

出　　处：《应用与环境生物学报》2011年第6期918-922,共5页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金项目(No.40801088);山东省自然科学基金项目(No.Q2008B09);山东省环境保护重点科技项目(No.2006007)资助~~

摘　　要：总RNA提取是进行实时荧光定量PCR、Northern杂交分析及cDNA文库建立等分子生物学实验研究的基础.提取出高质量高产量的土壤RNA已成为研究土壤微生物的关键步骤.综述了常用的几种土壤RNA提取方法,包括强变性剂法、Trizol法、CTAB法、SDS-Phenol法、酶解法以及试剂盒等方法.提取过程中化学试剂的选择及配比需依据不同的土壤类型,其中Trizol法在提取总RNA时应用广泛,再结合适当的物理破碎及生物学方法,可得到理想的RNA.试剂盒法提取土壤RNA方便快捷,纯度高但产量低,且对土壤具有选择性.最后提出了土壤RNA提取过程常见的一些问题和注意事项,为土壤RNA提取方法的选择提供依据.RNA plays an important role in studying environmental microbes because it can indicate the characteristics of the activated microorganism. The extraction of total RNA is the basis of real-time fluorescence quantitative PCR, northern hybridization analysis, cDNA library creation and other molecular biology studies. How to obtain a high yield and quality soil RNA is the critical step for the research of soil microbes. Therefore, several RNA isolation methods which can obtain high quality RNA from soil were summarized in this article, including strong denaturant method, Trizol method, CTAB method, SDS-phenol method, enzyme decomposition method and commercial RNA extraction kits. The various combinations of above methods for RNA isolation from soil were often used. Among the above methods, strong denaturant is good RNase inhibitor, and CTAB can cause the protein denaturation. However, bead beating is the efficient physical lysis method. Compared with other methods, mechanical homogenization in Trizol is the most efficient for releasing microbial RNA. The different RNA extraction kits have their characteristics for different soils. Furthermore, the influence factors, application aspects, as well as the potentials and the limitations for the extraction of total RNA were also reviewed. Therefore, this work may provide the guidelines for RNA isolation and the choice of method to study soil microbes. Ref 56

关 键 词：土壤微生物 RNA提取方法 裂解 RNASE 

分 类 号：S154.3[农业科学—土壤学]