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目的:研究牡丹皮基因组DNA提取的影响因素。方法:以根皮类中药材牡丹皮为材料,在快速少量抽提(CTAB)法的基础上对提取缓冲液中氯化钠、β-巯基乙醇浓度,水浴温度、时间,RNA酶(RNaseA)、聚合酶链(PCR)反应体系等条件进行考察。结果:使用经过改进后的CTAB法获得的DNA纯度和完整性较好,A260/A280值在1.8～2.0之间,PCR扩增条带清晰且亮,从而为接下来的分子生物学实验打下了良好的基础。结论:本方法经济、快速、高效,可作为根皮类中药材基因组DNA的提取方法,可为大规模生产提供理论依据。
Objective: To study the influencing factors of genomic DNA extraction from Cortex Moutan. METHODS: The root bark variety, Cortex Moutan, was used as the material. The concentrations of sodium chloride and β-mercaptoethanol in the extraction buffer, temperature and time of the water bath, and RNase A were determined on the basis of the CTAB method. The PCR reaction system and other conditions were investigated. Results: The purity and integrity of the DNA obtained with the improved CTAB method was better, the A260/A280 value was between 1.8 and 2.0, and the PCR amplification band was clear and bright, thus laying the foundation for the next molecular biology experiment. Good foundation. Conclusion: This method is economical, rapid, and efficient, and can be used as a method for extracting genomic DNA of root bark type Chinese herbal medicines, which can provide a theoretical basis for large-scale production.