改进的CTAB法提取中华芦荟叶总RNA

Extraction of Total RNA in Aloe vera L.var. chinensis Leaf with Improved CTAB Method

[目的]为分离中华芦荟次生代谢相关基因,构建成熟叶cDNA文库,探索从适合富含多糖和凝胶植物组织中提取高质量RNA的方法。[方法]以中华芦荟成熟叶为材料,采用改进的CTAB方法提取纯化芦荟叶总RNA。[结果]所提取的叶总RNAOD260/OD280值为1.88,电泳显示28 S1、8 S条带完整清晰,以此RNA为模板的RT-PCR结果显示目标特异条带。[结论]用该方法提取的RNA可用于cDNA合成、文库构建等后续分子生物学实验。

Objective The eDNA library of cloning genes related to secondary metabolites was constructed. Method The total RNA was extracted from mature Aloe vera L. var. chinensis leaf with polysaccharides and gel with improved CTBA method. Result The electrophoresis pattern showed the bands of 18 S rRNA and 28 S rRNA were sharp and complete, OD260/OD280 value of the extracted RNA was 1.88. The result of RT-PCR with the total RNA was positive. Conclusion These suggested the improved CTBA method could be suitable for the extraction of total RNA in Aloe vera L. var. chinensis with high quality and used in the subsequent molecular biological experiments such as reverse transcription and cDNA library construction.