从多糖丰富的样品中制备食用真菌DNA

Preparation of Edible Fungus DNA from Polysaccharide-Rich Sample

本文报道了一种从多糖丰富的食用真菌样品中快速、简便制备高分子量总DNA的改进方法。通过CTAB在不同的氯化钠浓度下选择沉淀DNA,除去多糖污染,苯酚、氯仿抽提,异丙醇沉淀,就可获得适宜多种分子生物学研究的食用菌基因纽DNA。本方法也适合小规模同时制备多个样品。另外,采用本方法制备的真菌DNA,经限制酶HaeⅢ酶切后在琼脂糖凝胶上能显示出菌株特异性的DNA带谱,可用于食用菌的遗传育种、菌株鉴定等研究。

A simple and efficient modified method for isolation of high molecular weight DNA from polysaccharide-rich edible fungus sample was reported. By the selection of precipitated DNA under different concentration of NaCl with CTAB to remove polysaccharide contamination, the extraction of crude DNA with phenol and chloroform and the precipitation of isopropanol, edible fungus genomic DNA pure enough for various molecular biological researches could be obtained. This method was also suitable to isolate many DNA samples at small scale simultaneouly. What of particular significance was that some strain-distinguished DNA band patterns could be seen on agarose gel after the total DNA was digested with restriction enzyme Hae Ⅲ. And this kind of Hae Ⅲ DNA bands could be used as a marker in the study of edible fungus strain breeding and identification.