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目的建立大黄根中总RNA提取的方法,为后续的分子生物学研究打下基础。方法以新鲜的大黄根为材料,分别采用Trizol法、常规CTAB-LiCl法、改良CTAB-LiCl法、通用型试剂盒法提取总RNA,利用琼脂糖凝胶电泳、微量紫外分光光度计和实时荧光定量PCR检测总RNA的纯度、完整性和产量。结果除CTAB-LiCl法外,Trizol法和通用型试剂盒法均不能从大黄根中提取出总RNA。同时,常规CTAB-LiCl法不稳定,所提总RNA有一定程度的降解。采用改良CTAB-LiCl法所提取的总RNA量大,纯度、完整性及质量均较好。结论改良CTAB-LiCl法对大黄等富含多酚、多糖类药材的总RNA提取具有一定的借鉴意义。
Objective To establish a method for the extraction of total RNA from Rhubarb roots and lay the foundation for subsequent molecular biology research. Methods Fresh rhubarb roots were used as materials. Trizol method, conventional CTAB-LiCl method, modified CTAB-LiCl method and universal kit were used to extract total RNA. The total RNA was extracted by agarose gel electrophoresis, micro-UV spectrophotometer and real- Quantitative PCR was used to measure the purity, integrity and yield of total RNA. Results In addition to CTAB-LiCl method, Trizol method and universal kit method can not extract total RNA from Rhubarb root. In the meantime, the conventional CTAB-LiCl method is unstable and the total RNA is degraded to a certain extent. The amount of total RNA extracted by modified CTAB-LiCl method is high, purity, integrity and quality are better. Conclusion The modified CTAB-LiCl method can be used as a reference for the extraction of total RNA from rhubarb and other polyphenols and polysaccharides.