高盐沉淀CTAB法提取温室菊花基因组DNA

Genomic DNA Extraction from Chrysanthemum Using High Salt CTAB Method

根据温室菊花植物组织富含多酚、多糖的具体特性,对CTAB法加以改进:在待沉淀液中加入1/2体积5 mol.L-1NaCl。改进后的方法获得的DNA质量良好,电泳条带清晰,提取过程无明显的DNA降解,基本上排除了多酚物质的干扰。以提取的DNA为模板,用一对引物扩增菊花中18S基因,得到条带单一,大小与已知一致,说明获得的DNA可以进行PCR扩增,EcoRI酶切基因组DNA图谱表明,提取的DNA能被限制性内切酶完全酶切,可以满足相关的分子生物学研究。

Based on CTAB method, some protocal was modified according to the physiological characteristics of high containing contents of polyphenolics and polysaccharides in Chrysanthemum. Adding 50% volume of 5mol·L^-1 NaCl2 into the solution which precipitating. High quality genomic DNA can be extracted from Chrysanthemum using the improved method. The DNA was preferable enough to be used as PCR template and digested by restriction enzyme completely and other studies of molecular biology.