富含多糖多酚的侧柏叶片总rna提取方法 - 吉林农业大学学报.pdfVIP

吉林农业大学学报 2011，11 Journal of Jilin Agricultural University E-mail:jlndxb@ 富含多糖多酚的侧柏叶片总RNA提取方法* 王暑辉，徐 倩，徐 筱，徐吉臣** 北京林业大学 生命科学与技术学院，北京 100083 摘 要：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 ；利用 SDS 酚法提取，提取物中含有大量多糖，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。 关键词：侧柏；RNA ；多糖；多酚；改进 SDS 酚法 中图分类号：Q522 ；Q949.667 文献标识码：A DOI: 网络出版地址： Extraction of Total RNA from the Leaves of Platycladus Orientalis Rich in Polysaccharides and Polyphenol WANG Shu-hui, XU Qian, XU Xiao, XU Ji-chen College of Life Science and Technology, Beijing Forestry University, Beijing 100083, China Abstract: 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. Key words: Platycladus orientalis ; RNA; polysaccharides; polyphenol; improved SDS-phenol method 侧柏（Platycladus orientalis ）又名扁柏，属柏科 作为