简便高质量的食用菌总RNA提取方法  

A Rapid and Efficient Method for Isolating High Quality Total RNA from Edible Fungi

作　　者：税丕容[1] 郑晓冰[1] 林俊芳[1] 郭丽琼[1] 

机构地区：[1]华南农业大学食品学院生物工程系,广东广州510640

出　　处：《食用菌学报》2008年第1期32-41,共10页Acta Edulis Fungi

基　　金：国家自然科学基金(编号:30671457;30371000);广东省自然科学基金(编号:5006680;032239)的部分研究内容

摘　　要：STE法是针对食用菌多糖含量高而设计的一种提取食用菌总RNA的有效方法。本研究以灵芝、香菇和白腐真菌TR16菌株的菌丝体为材料,采用STE法提取其总RNA,同时以Trizol法作对照。结果表明:采用STE法提取的三种菌丝的总RNAA260/A280比值在1.80～2.10范围内,A260/A230略大于2.00;而Trizol法提取的总RNAA260/A280比值小于1.80,A260/A230小于1.90,明显比STE法效果差;总RNA电泳检测表明:STE法的3条带(28SrRNA、18SrRNA和5SrRNA)清晰,无拖尾,无杂带,效果比Trizol法好;将STE法提取的总RNA进行反转录后合成双链cDNA,进行预扩增,电泳显示呈弥散状;再进行选择性扩增,有相同或相异的条带。进一步表明STE法提取的总RNA质量较高,可以满足后续分子生物学研究的要求。A rapid and efficient procedure, the STE （sodium chloride-Tris-HCI-EDTA） method, for isolating high quality total RNA from polysaccharide-rich mycelia of the edible fungi Lentinus edodes, Ganoderma lucidum and a Polyporus sp. is described. The quality of the total RNA isolated by the STE method was superior to that obtained using the Trizol method. The A260/A280 and A260/A230 ratios of RNA isolated by the STE method ranged between 1.80-2.10 and between 1.90-2.30 respectively, whereas the A260/A280 ratio of RNA isolated by the Trizol method was below 1.80, and the A260/A280 ratio was outside the 1.90-2.30 range. Three distinct bands representing 28S, 18S and 5S rRNA were observed following electrophoresis of total RNA isolated from the three test strains by the STE method. Electrophoresis of total RNA isolated using the Trizol method resulted in dark bands corresponding to 28S rRNA and 18S rRNA, indicating partial degradation of the RNA. Our data demonstrated that the total RNA isolated by the STE method was of a quality eminently suited for downstream applications.

关 键 词：食用菌 总RNA STE法 

分 类 号：S646[农业科学—蔬菜学]