3种飞机草总RNA提取方法比较研究

Extraction methods of total RNA from Eupatoriumm odoratum L.

以飞机草幼嫩叶片为材料,分别采用Trizol法、异硫氰酸胍法、CTAB法对叶片总RNA进行提取。结果表明,异硫氰酸胍法和Trizol法提取的飞机草叶片总RNA质量较高,以异硫氰酸胍法提取的RNA浓度和纯度更优,蛋白质污染少,可用于基因克隆;CTAB法不能提取到RNA或提取的RNA含量极低,电泳检测不到任何条带,不适于提取飞机草总RNA。因此,异硫氰酸胍法和Trizol法可用于研究飞机草入侵适应性的分子机理。

In the present study, three methods including Trizol, guanidine isothiocyanate and CTAB were used to extract total RNA from young leaves of Eupatoriumm odoratum L. The results showed that the quality of total RNA extracted by Trizol and guanidine isothiocyanate method was better than that of CTAB, but the concentration and purity of RNA extracted by guanidine isothiocyanate method were better, which could be used for gene cloning. The concentration and purity of RNA isolated by CTAB method was poor and the bands could not be detected after RT-PCR ＆ amplification by PCR. Therefore, CTAB method was unsuitable for extracting the total RNA from leaves of Eupatoriumm odoratum L., and Trizol and guanidine isothiocyanate methods could be utilized to study the molecular mechanism on invading adaptability of Eupatoriumm odoratum L.