苎麻总DNA提取的CTAB法优化方案  

Optimized CTAB Protocol for Extracting the Total DNA of Ramie

作　　者：侯思名[1] 段继强[1] 梁雪妮[2] 丁小维[1] 肖伏[1] 刘飞虎[1] 

机构地区：[1]云南大学生命科学学院,昆明650091 [2]云南大学成人教育学院,昆明650091

出　　处：《西北植物学报》2005年第11期2193-2197,共5页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金资助项目(30360058)

摘　　要：根据苎麻组织自身含有较多的酚类、粘状物及黄酮类等次生代谢物质的特点,对CTAB法进行优化后用于提取苎麻组织总DNA.在研磨过程中加入抗氧化剂PVP,提取缓冲液中加入终浓度6%的PVP和4%的β-巯基乙醇,可防止整个提取过程中溶液变褐;在第2次用氯仿/异戊醇抽提时,加入10%的CTAB和4%的N aC l高盐溶液,以除去多糖.经几种改良方案提取总DNA的ISSR-PCR扩增结果验证,以同时加入抗氧化剂和高盐溶液的CTAB-4法效果最好.Considering that ramie is rich in secondary metabolites such polyphenols, sticky substances and flavone, the optimized CTAB protocol was screened for extracting the total DNA of ramie. The addition of polyvinylpyrrolidone（PVP） during the grinding and the additions of Polyvinylpyrrolidone at the final concentration of 6% and β-mercaptoethanol at the final concentrations of 4% to the extraction buffer were capable of preventing browning from taking place in the extraction; while chloroform/ isoamyl alcohol were used to conduct the second extraction, 10% CTAB and 4% NaCI were added to remove polysaccharides. The total DNAs extracted by the several modified CTAB protocols were amplified by ISSR-PCR and the amplification proved that CTAB-4 in which both the antioxidant and the salt at high concentration were added performed best.

关 键 词：苎麻 DNA提取 CTAB法 优化 

分 类 号：Q78[生物学—分子生物学] S563.1[农业科学—作物学]