摘要
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 for the extraction of the genomic DNA from Paeonia Suffruticosa. METHODS： Taking Paeonia Suffruticosa （root bark of Chinese medicinal herb） as material to investigate the influencing factors including concentrations of the NaCt and beta- mercaptoethanol, temperature and time of water bath, RNaseA, PCR （polymerase chain reaction） system etc in the buffer solution on the basis of modified CTAB method. RESULTS： The DNA obtained by modified CTAB method was pure, integrated, with the value of A260/A280 ranged from 1.8 to 2.0, the amplified bands of PCR were clear and bright, which lay a solid foundation for the following molecular biology experiments. CONCLUSION： The modified CTAB method is economical, rapid and efficient, and it can be served as an extraction of genomic DNA from root bark Chinese medicinal herb as well as a theoretical basis for full scale production.
出处
《中国药房》
CAS
CSCD
北大核心
2008年第27期2084-2087,共4页
China Pharmacy
基金
重庆市自然科学基金资助项目(7351)
