克隆植物珠芽蓼基因组DNA的提取及RAPD鉴定

高原植物体内所含有的酚类、多糖、色素等次生代谢物质严重影响提取其基因组DNA的得率和纯度,是导致后续分子操作失败的主要原因.运用一种新改进的CTAB法对变色硅胶保存的高原植物珠芽蓼(Polygonumviviparum)叶片进行基因组DNA的提取,采取常温下沉淀DNA,增加洗涤沉淀的体积分数为70%乙醇用量和洗涤时间等措施,有效地去除了酚类、多糖、色素等物质的干扰,减少了褐化现象的发生.样品检测所得DNA片段均在48 kb左右,电泳谱带呈线状,无拖尾现象;紫外分光光度计检测,A260nm/A280nm值在1.7~1.9之间.RAPD扩增结果表明,此方法提取的DNA无论在质量和纯度上都较理想.

Extraction of genomic DNA from Polygonum viviparum and its RAPD analysis

Many compounds,such as polysaccharides,coloring matters,phenolic compounds and other secondary metabolites,in the alpine-plants influence on the yield and quality of genomic DNA while it is extracted and that lead up to the failure of following molecular operation.This study developes an improved CTAB method to extract the genomic DNA in leaves(which are dried by silica gel)of Polygonum viviparum.By taking some measures,such as precipitation at room temperature,increasing the volume of alcohol to wash the precipitated DNA,the phenolic compounds,polysaccharides and coloring matters are removed effectively.The size of the obtained DNA is about 48 kb and the value of A_(260nm)/A_(280nm) isolated DNA is between 1.7~1.9.The result of RAPD indicates that the quantity and quality of DNA got by the improved CTAB method are all ideal.