荒漠生物结皮中齿肋赤藓总RNA最佳提取方法的确立  

A Method Established for Total RNA Extraction from Syntrichia caninervis in a Temperate Desert

作　　者：马瑞[1] 杨红兰[2] 张元明[2] 张道远[2] 徐刚标[1] 

机构地区：[1]中南林业科技大学生命科学院,湖南长沙410004 [2]中国科学院干旱区生物地理与生物资源重点实验室中国科学院新疆生态与地理研究所,新疆乌鲁木齐830011

出　　处：《生物技术》2010年第3期52-54,共3页Biotechnology

基　　金：国家重点基础研究发展计划项目(2009CB825104);新疆自治区重大专项(200731138-3);中科院知识创新工程(KZCX2-YW-336)资助

摘　　要：目的:根据不同提取方法对提取荒漠齿肋赤藓总RNA质量的影响,寻找一种快速高效提取齿肋赤藓总RNA的方法。方法:采用改良RNAiso Reagent试剂法、RNAprep pure植物总RNA提取试剂盒法(TIANGEN)、皂土法分别提取采自新疆古尔班通古特沙漠生物结皮中齿肋赤藓的总RNA,用紫外分光光度计测定其浓度、纯度和产量,并通过RT-PCR进一步检测其质量。结果:三种方法均能提取出RNA,但质量差异较大。其中改良的RNAiso Reagent试剂法提取的RNA产量可达1298～1318μg/g,完整性好且纯度高,OD260/OD280值为1.86～1.92,且经济、操作简便,能有效抑制多酚物质氧化和次生代谢物质的影响,,适合用于进一步的RT-PCR反应。结论:改良的RNAisoReagent法经济且操作简便,提取的荒漠齿肋赤藓RNA完整性和纯度较高,可用于RT-PCR反应。Objective：To determine a rapid and efficient method for extracting total RNA from Syntrichia caninervis,a drought tolerant desert moss,three tridational methods were selected and examined.Method：Methods of modified RNAiso Reagent,bentonite and RNAprep pure total RNA isolation kit（TIANGEN） were chose to extract RNA from S.caninervis.The purity,quantity and productivity of extracted RNA were determined by UV spectrophotometer,and its quality was measured through RT-PCR.Result：The qualities of total RNA isolated from S.caninervis through three methods respectively were significantly different.Among three tested methods,the modified RNAiso Reagent method could present an optimal result,and efficiently inhibited the influence of phenolic compounds and other secondary substances.The productivity of total RNA extracted by this method could reach 1 298-1 318μg/g,and the ratio of OD260 /OD280 could be 1.86 to 1.92.Conclusion：The modified RNAiso Reagent is a simple and convenient way for total RNA extraction,and could provide extracted RNA with good quality and high purity which is suitable for further molecular research such as RT-PCR.

关 键 词：齿肋赤藓 总RNA 提取 改良RNAiso Reagent法 古尔班通古特沙漠 

分 类 号：Q781[生物学—分子生物学]