Ding Ling-Wen, Sun Qiao-Yang, Wang Zhao-Yu, Sun Yong-Bin, Xu Zeng-Fu
Key Laboratory of Gene Engineering of the Ministry of Education and State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
Anal Biochem. 2008 Mar 15;374(2):426-8. doi: 10.1016/j.ab.2007.11.030. Epub 2007 Nov 28.
The most commonly used protocol of the RNA isolation, the guanidine thiocyanate method, was unsuitable for recalcitrant plant tissues containing a large amount of storage proteins and secondary metabolites. We demonstrated that RNA could bind to the silica particles, which have been used successfully in DNA isolation from various sources, under a high concentration of NaCl in the presence of ethanol and sodium acetate. Based on this observation, an efficient, inexpensive, and highly reproducible technique, the acid phenol-silica method, was developed to isolate high-quality RNAs from various plant tissues recalcitrant to extraction in guanidine thiocyanate.
最常用的RNA提取方法——硫氰酸胍法，不适用于含有大量贮藏蛋白和次生代谢物的难提取植物组织。我们证明，在乙醇和醋酸钠存在的高浓度氯化钠条件下，RNA能够与二氧化硅颗粒结合，而二氧化硅颗粒已成功用于从各种来源提取DNA。基于这一观察结果，我们开发了一种高效、廉价且可高度重复的技术——酸性酚-二氧化硅法，用于从硫氰酸胍提取困难的各种植物组织中分离高质量的RNA。
