摘要
从植物组织中提取高质量的RNA是进行cDNA文库构建等分子生物学研究的前提。在苯酚法的基础上,改进并得到了一种适合紫茎泽兰根、茎、叶总RNA快速提取的方法,消除了蛋白质、DNA、多糖等的污染。该方法提取的紫茎泽兰不同组织总RNA纯度高、完整性好,可用于RT-PCR、cDNA文库构建、Northern杂交等分子生物学实验,而且简单、经济、重复性好,适合于多种植物组织总RNA的提取。Northern杂交表明F3’H基因在紫茎泽兰的根、茎、叶等组织中广泛存在,但在叶中的表达量最高,在根中的表达量最低。
Extracting high quality RNA from plant tissues is the groundwork in carrying out researches in molecular biology such as cDNA library construction, In this study,an improved method based on the PHENOL extraction method were established to extract total RNA from roots,stems and leaves of E. aclenophorum effuziently. It approved to be able to avoid protein,DNA and polysaccharide contaminations. And the total RNA extracted from different E. adenophorum tissues was of high purity and integrity,and was suitable for RT-PCR,northern blot and cDNA library construction. This method is simple,economic,quick and has good repeatability,and is also suitable for extracting total RNA from other plant species. Northern blot analysis showed that the F3＇H gene expressed in the leaves,stems and roots of E. adenophorum,and the highest expression level of F3＇H was detected in the leaf, then the medium in the stem,and the lowest in the root.
出处
《生物技术通报》
CAS
CSCD
2007年第2期147-150,共4页
Biotechnology Bulletin
