苜蓿总RNA提取方法比较研究  

Comparative Study on of Extraction Methods of Total RNA from Medicago sativa

作　　者：何庆元[1,2] 吴萍[1] 李正鹏[1] 张晓红[1] 胡琴[1] 

机构地区：[1]安徽科技学院生命科学学院,安徽凤阳233100 [2]南京农业大学大豆研究所／国家大豆改良中心／作物遗传与种质创新国家重点实验室,江苏南京210095

出　　处：《中国草地学报》2009年第1期116-120,共5页Chinese Journal of Grassland

基　　金：国家“973”计划基金项目(2007CB108901);安徽省教育厅基金项目(2006KJ205B)

摘　　要：以苜蓿叶片为材料,利用紫外分光光度计和凝胶电泳法比较了改良CTAB法、异硫氰酸胍法、SDS法、Tris-热硼法和尿素法等五种方法提取的苜蓿总RNA的质量和纯度。结果表明:改良CTAB法提取的RNA的OD260nm/OD230nm大于2.0并且OD260nm/OD280nm接近1.8,具有较高的纯度。凝胶电泳结果表明,改良CTAB法有28SrRNA和18S rRNA两条清晰的条带,且无降解;其它四种方法获得的RNA品质较差,有降解和弥散现象。将改良CTAB法提取的RNA逆转录成cDNA,经RAPD扩增后出现4条清晰的条带,进一步证明了改良CTAB法提取的总RNA具有很高的纯度,可以满足进一步分子生物学研究的要求。Modified CTAB method, Guanidine Thiocyanate Method, SDS Method, Tris-- H3BO4 Method and Urea Method were employed to extract total RNA in leaf of Medicago sativa. The quality and quantity of total RNA were compared among five methods through UV spectrometer and gel electrophoresis. The results indicated that the value of OD260nm/OD230nm of RNA extracted with modified CTAB method were greater than 2.0 and the value of OD260nm/OD280nm were approach to 1.8, which showed higher pourity. Gel electrophoresis showed that RNA extracted with modified CTAB method had clear bands of 28S rRNA and 18S rRNA and was not degraded; RNA extracted with other four methods was degraded and dispersed in some degrees. RNA extracted with modified CTAB method could be reversed to eDNA. The eDNA was used for RAPD amplify and four clear bands could be observed in agarose gel. These results further demonstrated that the quality and purity of the total RNA extracted with modified CTAB method can meet the demands of molecular biology experiment.

关 键 词：苜蓿 总RNA 提取方法 

分 类 号：Q503[生物学—生物化学]