【摘要】 研究了一种可有效排除茶树叶片细胞所富含的茶多酚、茶多糖等杂质污染的茶树总RNA提取方法.即在Trizol法的基础上,于提取的初始阶段,重点防止褐化效应,用水不溶性聚乙烯吡咯烷酮(PVP)和β-巯基乙醇(β-Me)排除茶多酚干扰,同时用低含量乙醇去除茶多糖;在最后阶段,55-60℃热水浴10min,低温离心进一步去除茶多糖的干扰.利用这一方法提取的茶树总RNA,经琼脂糖电泳鉴定,可见清晰的18s和28sRNA的2条带型,证明RNA完整,无弥散或降解.更多还原

【Abstract】 A method of isolating RNA from tea plant was studied. It could effectively remove polyphenols and polysaccharides, which are rich in tea leaf cells. The modification based on the extraction procedure by Trizol, tea polyphenols were removed with water insoluble polyvinylpyrrolidone (PVP) and βMercaptoethanol (βMe) for fear of browning effect at the beginning stage of isolation. Tea polysaccharides were also dealt with lowconcentration alcohol at the same time. The polysaccharides were further removed by incubation for 10 min (55-60 ℃), followed by centrifugation at 4 ℃ in the end. With the modified method, two clear bands of 18 s RNA and 28 s RNA isolated from tea plant could be found by agarose gel electrophoresis. It was proved that RNA was intact, without smear or degradation.更多还原