曾凡亞; 張義正
四川大學生物工程系; 四川大學生物工程系 成都 610064 ; 成都 610064
【中文摘要】 本文報道了一種從多糖豐富的食用真菌樣品中快速、簡便制備高分子量總DNA的改進方法。通過CTAB在不同的氯化鈉濃度下選擇沉淀DNA,除去多糖污染,苯酚、氯仿抽提,異丙醇沉淀,就可獲得適宜多種分子生物學研究的食用菌基因紐DNA。本方法也適合小規模同時制備多個樣品。另外,采用本方法制備的真菌DNA,經限制酶HaeⅢ酶切后在瓊脂糖凝膠上能顯示出菌株特異性的DNA帶譜,可用于食用菌的遺傳育種、菌株鑒定等研究。
【英文摘要】 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.
【中文關鍵詞】 DNA制備; 食用菌; 多糖
【英文關鍵詞】 DNA isolation; Edible fungus; Polysaccharide
【文獻出處】 食用菌學報,Acta Edulis Fungi,編輯部郵箱,1996年03期 【DOI】CNKI:SUN:SYJB.0.1996-03-002