木麻黄小枝总RNA提取方法的比较与优化  

Comparison and optimization of total RNA extraction methods from twigs of Casuarinaspp.

作　　者：孙思[1,2] 陈洁玲[1] 舒灿伟[1] 王军[2] 王新荣[1] 周而勋[1] 

机构地区：[1]华南农业大学资源环境学院,广州510642 [2]华南农业大学林学院,广州510642

出　　处：《华中农业大学学报》2014年第1期51-55,共5页Journal of Huazhong Agricultural University

基　　金：国家科技支撑计划课题"沿海防护林主要病虫害防控技术研究"(2009BADB2B0203-01)

摘　　要：分别采用Trizol法、CTAB法、异硫氰酸胍法、SDS法和Omega试剂盒法从短枝木麻黄小枝中提取总RNA。结果表明:用Trizol法和异硫氰酸胍法没有提取到木麻黄总RNA;用CTAB法提取的RNA发生了降解;只有SDS法和Omega试剂盒法成功提取到木麻黄小枝RNA,但质量均没有达到试验要求。对提取效果相对较好的SDS法进行改进,主要是加入去除次生代谢物质的步骤。经检测,改进后的SDS法提取的木麻黄小枝总RNA纯度高、完整性好,能够满足RT-PCR分析等分子生物学研究的要求。High quality RNA extracted from the twigs of Casuarina equisetifolia is used as the ba- sis for the study of its gene expression. Five RNA extraction methods, i. e. Trizol method, CTAB method, guanidine thiocyanate method, SDS method and Omega RNA isolation kit, were used to ex- tract total RNA from the twigs of C. equisetifolia. The results showed that RNA was unable to be ex- tracted with Trizol and guanidine thiocyanate methods, and the RNA extracted with CTAB method was degraded; only SDS and Omega RNA isolation kit methods could succeessfully to extract RNA from the twigs of C. equisetifolia, however, the RNA quality didnrt meet the requirement of further experiment. SDS method, the most favourable methods in this study, was improved to get better results, and the major improvement was the addition of steps for removing secondary metabolites. Evaluation experi ment indicated that RNA with high quality and yield was obtained from the improved SDS method, and could be used for further study of molecular biology such as RTPCR.

关 键 词：木麻黄 小枝 总RNA 提取方法 

分 类 号：S763[农业科学—森林保护学] S792.93[农业科学—林学]