丹参基因组DNA提取方法比较研究
Comparative study on genomic DNA extraction methods of Salvia miltiorrhiza
李娟 1姜鑫萍 1许张雨 1张贵林 1杨奕樱1
作者信息
- 1. 贵州中医药大学,贵州 贵阳 550025
- 折叠
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.
Abstract
In this study we investigated the best method for obtaining genomic DNA from Salvia miltiorrhiza.Ge-nomic DNA was extracted from Salvia miltiorrhiza leaves by different methods.The integrity was evaluated via gel e-lectrophoresis,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 ob-tained by the modified urea method had poor amplification effect;β-actin could be amplified with the DNA extrac-ted by the 1.5×CTAB method,the 2×CTAB method and the high salt-low pH method.In conclusion,the DNA ex-tracted 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/CTABKey words
Salvia miltiorrhiza/genomic DNA extraction/method comparison/CTAB引用本文复制引用
基金项目
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)
出版年
2024
NETL
NSTL
万方数据