摘要
针对药用植物丹参含有较多的多酚类、醌类等水溶性次生代谢物,在提取DNA时易发生共沉淀而很难除去等问题,设计双蒸水预洗不同次数和活性碳吸附杂质等处理,对CTAB和SDS两种DNA提取方法进行了系统优化。研究得出,采用2%的CTAB提取液和双蒸水预洗一遍并且添加样品鲜重4%的活性碳于提取液中的提取方法最佳。不仅能除去多酚类、醌类等次生代谢物,而且去多糖、蛋白质效果良好,所获DNA纯度高;RAPD-PCR实验结果稳定清晰可靠。
Salvia Miltiorrhiza Bge contains a plenty of secondary metabolites, such as polyphenol derivatives and quinonoid compounds which could combine DNA to form sediments and are hard to be removed. On the basis of optimization for pre-wash times with redistilled water and proportions of charcoal added, two DNA-extraction methods of CTAB and SDS were compared. The results showed that the method （2% CTAB solution, pre-wash once and 4% of weight of fresh samples for charcoal） was optimal. This method has the advantages that not only polyphenol and quinonoid compounds, but also polysaccharide and proteins are eliminated. As a result, high purity DNA was obtained to be used as the model of RAPD-PCR and the electrophoretogram of DNA amplification product was clear and credible in this research.
出处
《药物生物技术》
CAS
CSCD
2007年第1期29-33,共5页
Pharmaceutical Biotechnology
基金
教育部新世纪优秀人才支持计划
西北农林科技大学拔尖人才支持项目
关键词
丹参
次生代谢物
CTAB
双蒸水预洗
活性炭
Salvia miltlorrhiza Bge, Secondary metabolites, CTAB, Pre-wash with double distilled water, Activated charcoal
