富含多糖多酚的侧柏叶片总RNA提取方法

以侧柏叶片为试验材料,利用Trizol法、CTAB法、SDS酚法和改进的SDS酚法提取RNA,结果表明:Trizol法和CTAB法都不能提出侧柏叶片总RNA；SDS酚法提取,提取物中含有大量多糖,提取的RNA质量少,不足以用于后续的研究；改进的SDS酚法提取的RNA纯度高,电泳效果好.进一步通过反转录和特异引物扩增试验发现:利用改进的SDS酚法提取的RNA反转录效果好,能扩增出预期的基因片段,可很好地应用于基因克隆与基因表达分析等分子生物学试验研究.

Extraction of Total RNA from the Leaves of Platycladus orientalis Rich in Polysaccharides and Polyphenol

The study compared the four methods of Trizol,CTAB,SDS-phenol,and improved SDS-phenol in extraction of total RNA from the leaves of Platycladus orientalis.The electrophoresis results showed that Trizol and CTAB method could not be successfully used for RNA isolation from Platycladus orientalis leaves.SDS-phenol method could be used for isolating RNA,but with abundant polysaccharides inside it did not satisfy the further experiments.Comparably,the improved SDS-phenol method could extract RNA from Platycladus orientalis leaves with higher quality through adding low concentration of ethanol and others.The further experiments displayed that the total RNA could be well transcribed and used for the specific amplification of gene fragments.It confirmed that the total RNA isolated by the improved SDS-phenol method could be well used for future gene cloning and expression studies.