响应面法优化海南野牡丹属植物DNA的提取条件

Optimization of DNA Extraction Condition of Hainan Melastoma L. Based on Response Surface Methodology

为研究不同环境中的野牡丹是否为同一生态型,分析海南野牡丹的种间关系。本研究建立、优化了海南省野牡丹属植物DNA提取条件,利用DX8Trial软件分别对CTAB浓度、PVP浓度、β-巯基乙醇浓度、醋酸钠用量等因素进行改良设计。结果表明,最佳提取条件为:2%CTAB、2%PVP、1%β-巯基乙醇、5 mol/L醋酸钠90μL;同时,采用该条件提取野牡丹属其它植物DNA,结果 DNA纯度较高,条带清晰,提取效果好。最终该提取条件适用于海南产野牡丹属植物DNA的提取,为该属植物遗传多样性研究及分子生物学研究提供依据。

In order to study whether Melastoma L. in different environments were the same ecotypes, and analyze the interspecific relationship of Hainan Melastoma, the study established and optimized the DNA extraction conditions of Melastoma in Hainan Province, and used the DX8 Trial software to improve the CTAB concentration,PVP concentration, β-mercaptoethanol concentration, sodium acetate content and other factors. The results showed that the optimum extraction conditions were as follows： 2% CTAB, 2% PVP, 1% β-mercaptoethanol and 5 mol/L sodium acetate 90 μL. Meanwhile, using this condition to extract the DNA of other plants in Melastoma,the result showed higher DNA purity and clear bands as well as good extraction effect. At last, the extraction condition was applicable to the extraction of the plant DNA of Hainan Melastoma, and might provide a basis for the study of genetic diversity of Melastoma plant, and molecular pharmacognosy.