皖贝母总RNA提取方法的比较研究

Comparison of different methods for isolating total RNA from bulblet of Fritillaria anhuiensis

目的:针对皖贝母鳞茎富含多糖、酚类和其他多种次生代谢产物的特点,筛选一种简单高效的皖贝母总RNA提取方法。方法:用异硫氰酸胍法、皂土法、改进的SDS/酚法、RNAiso plus 4种方法分别提取皖贝母鳞茎总RNA,并对提取的RNA进行电泳及RT-PCR技术检测与分析。结果:皂土法提取的总RNA条带清晰无弥散,完整性好无明显DNA和其他杂质污染,适用于RT-PCR试验。结论:皂土提取法是一种适合于皖贝母总RNA抽提的简单高效的方法。

Objective： To optimize a simple and effective method for total RNA extraction from bulblet of Fritillaria anhuiensis. Method： Four methods, i.e. guanidine isothiocyanate, bentonite, modified SDS/phenol and the RNAiso plus, were used to extract total RNA from bulblet of F. anhuiensis. Then the results of the extraction were compared and analyzed by electrophoresis detection and RT-PCR verification. Result： The total RNA extracted by bentonite method were clear and no dispersion, the integrity of the RNA was well, and there was no obvious contamination with DNA and other impurities, was suitable for RT-PCR test. Conclusion： The bentonite method is quick, economic, and efficient for total RNA extraction from bulblet of F. anhuiensis.