丹参基因组DNA提取方法比较研究  

Comparative study on genomic DNA extraction methods of Salvia miltiorrhiza

作　　者：李娟 姜鑫萍 许张雨 张贵林 杨奕樱 LI Juan;JIANG Xinping;XU Zhangyu;ZHANG Guilin;YANG Yiying(Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China)

机构地区：[1]贵州中医药大学,贵州贵阳550025

出　　处：《贵州科学》2024年第3期31-35,共5页Guizhou Science

基　　金：贵州中医药管理局中医药、民族医药科学技术项目(QZYY-2022-013);贵州省教育厅青年科技人才成长项目(黔教合KY〔2021〕193);国家自然科学基金地区基金(82160719);贵州中医药大学博士启动基金(〔2020〕30)。

摘　　要：目的:为了探究最适合丹参基因组DNA的提取方法。方法:本研究以丹参叶片为材料,采用不同的方法提取其基因组DNA,通过凝胶电泳检测其完整度、微型分光光度计检测浓度及纯度、看家基因扩增等进行比较,以选择出最适宜的提取方法。结果:应用碱裂解煮沸法、尿素法不能得到完整DNA;4×CTAB法所得的DNA有RNA污染;SDS冰浴法、PVP-40法和尿素法提取的DNA有蛋白质和酚类污染;改良尿素法所得DNA的看家基因扩增效果不好;1.5×CTAB法、2×CTAB法和高盐低pH法提取的DNA能保证看家基因的扩增。结论:2×CTAB法提取的DNA浓度和纯度高,可用于后续相关分子生物学研究。In this study we investigated the best method for obtaining genomic DNA from Salvia miltiorrhiza.Genomic DNA was extracted from Salvia miltiorrhiza leaves by different methods.The integrity was evaluated via gel electrophoresis,the concentration and purity were assessed with a micro-spectrophotometer,and the amplification of β-actin was compared.The results showed that the alkali pyrolysis boiling method and the urea method could not provide full length DNA;RNA was present in the DNA extracted by the 4×CTAB method;proteins and phenol were present in the DNA acquired by the SDS ice bath method,the PVP-40 method and the urea method;the DNA obtained by the modified urea method had poor amplification effect;β-actin could be amplified with the DNA extracted by the 1.5×CTAB method,the 2×CTAB method and the high salt-low pH method.In conclusion,the DNA extracted by the 2×CTAB method had high concentration and purity,and was suitable for use in subsequent molecular biology studies.

关 键 词：丹参 基因组DNA提取 方法比较 CTAB 

分 类 号：S567.2[农业科学—中草药栽培]