一种高效经济的高质量植物RNA提取方法  

An Efficient and Economic Method for Preparation of High Quality Plant RNA

作　　者：张年辉[1] 韦振泉[1] 何军贤[1] 杜林方[1] 梁厚果[1] 

机构地区：[1]四川大学生命科学学院,成都610064

出　　处：《生物化学与生物物理进展》2004年第10期947-950,共4页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金资助项目 ( 3 0 2 70 12 1);国家教育部博士点基金资助项目 ( 2 0 0 2 0 610 0 94)~~

摘　　要：建立了一种高效经济的植物RNA提取方法 .在提取缓冲液中加入蔗糖、氯化钾和镁离子以提供对RNA分子的保护 .破碎后的细胞于提取缓冲液中裂解后 ,用酚 /氯仿变性并去除内源RNA酶和其他蛋白质 ,而后用pH 5 6 的NaAc沉淀RNA .用该方法提取RNA的得率较高 ,经电泳检测 ,RNA的完整性很好 .RNA印迹分析和RT PCR也都得到很好的结果 .该方法还使实验成本大大降低 .An efficient and economic method for high quality RNA preparation from plant tissues was established. To avoid RNA degradation, sucrose, potassium chloride and Mg2+ were included in the extraction buffer. Plant tissues were lysed in the extraction buffer, then ribonucleases (RNAses) and other proteins were denatured and extracted by phenol/chloroform. After that, the DNA was selectively fractionated from RNA with sodium acetate (NaAc) (pH 5.6). The isolated RNA with this method gave good yield. Results of non-denaturing electrophoresis or formaldehyde agarose gel electrophoresis both showed higher amount of 25 S ribosomal RNA (rRNA) than that of 18 S rRNA. Northern hybridization gave sharp and clear signals. Both plastid gene and nuclear gene were amplified successfully by RT-PCR. These results show that, the RNAs isolated with this method are in good integrity and purity, and can meet the needs of most molecular biological experiments including gene cloning and expression analysis. In this method, phenol/chloroform were used to remove proteins and inactivate RNAses, NaAc (pH 5.6) was used to precipitate RNAs, thus largely reduced the experimental expenses.

关 键 词：RNA分离 植物 RNA酶 

分 类 号：Q781[生物学—分子生物学]