喜马拉雅紫茉莉不同组织总RNA提取方法的比较分析  

Comparative Analysis of Extraction Methods of Total RNA from Different Tissues of Mirabilis himalaica

作　　者：袁芳 王靖 王昭凤 兰小中 YUAN Fang;WANG Jing;WANG Zhaofeng;LAN Xiaozhong(Food Science College,Tibet Agriculture&Animal Husbandry University,Nyingchi Tibet,860000;Tibetan Collaborative Innovation Center of Agricultural&Animal HusbandryResources,Food Science College,Tibet Agriculture&Animal Husbandry University,NyingchiTibet,860000,China)

机构地区：[1]西藏农牧学院食品科学学院 [2]西藏农牧学院食品科学学院西藏特色农牧资源协同创新中心,西藏林芝860000

出　　处：《高原农业》2021年第5期485-489,523,共6页Journal of Plateau Agriculture

基　　金：林芝市科技计划项目—校地合作项目(XDHZ-2020-01);西藏自治区科技计划项目科技重大专项(XZ201901-GA-04)。

摘　　要：为了从喜马拉雅紫茉莉不同组织中提取高质量的总RNA,本研究以喜马拉雅紫茉莉的根、茎和叶组织为材料,采用Trizol法、改良Trizol法、CTAB-LiCl法和试剂盒法提取总RNA,使用琼脂糖凝胶电泳和超微量紫外分光光度计检测RNA的完整性、纯度与浓度。结果表明:Trizol法可用于叶组织RNA的提取,但RNA纯度低;改良Trizol法可用于茎和叶组织RNA的提取,RNA完整性较好,有少量的蛋白质、酚类物质和有机溶剂残留,茎和叶组织RNA的浓度分别为385.43 ng/μL、394.54 ng/μL;CTAB-LiCl法提取的根、茎和叶组织总RNA的浓度较低,并且RNA纯度差,A260/A280比值均大于2.0,A260/A230比值均小于2.0,RNA发生严重降解;试剂盒法提取的根、茎和叶组织总RNA完整性好,浓度高分别为:451.92 ng/μL、579.35 ng/μL和664.97 ng/μL,并且RNA纯度高,A260/A280比值均在1.8~2.0,A260/A230比值均大于2.0。因此,Trizol法和CTAB-LiCl法不适用于提取喜马拉雅紫茉莉总RNA,而改良Trizol法和试剂盒法可提取到质量较好的喜马拉雅紫茉莉总RNA。In order to extract high quality total RNA from different tissues of Mirabilis himalaica,the four RNA extraction methods,such as Trizol method,Trizol improved method,CTAB-LiCl method and kit method were performed using the young leaves,tender stems,and roots of M.himalaica,and the integrity of RNA were tested by agarose gel electrophoresis,whereas the purity and concentration of total RNA were detected by ultramicro UV spectrophotometer.The results showed that the Trizol method could be used to extract RNA from leaf tissue,but the purity of RNA were low.The Trizol improved method could be used to extract RNA from stem and leaf tissues,the RNA integrity were good,and a small amount of protein,phenolic substances and organic solvents remained,the concentration of RNA were 385.43 ng/μL and 394.54 ng/μL,respectively.The RNA concentration and purity extracted by CTAB-LiCl method were low,and the RNA degradation were serious.The kit method had the best effects of extracting total RNA from the root,stem and leaf tissue of M.himalaica,with the highest concentrations of 451.92 ng/μL,579.35 ng/μL and 664.97 ng/μL,respectively,and their A260/A280 were both within the range of 1.8~2.0,while A260/A230 were both higher than 2.0,showing high purity.Therefore,Trizol method and CTAB-LiCl method were not suitable for extracting total RNA from M.himalaica,while Trizol improved method and kit method could extract better quality total RNA from M.himalaica.

关 键 词：喜马拉雅紫茉莉 总RNA 提取方法 

分 类 号：Q522[生物学—生物化学]