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.
出处
《华中农业大学学报》
CAS
CSCD
北大核心
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)
关键词
木麻黄
小枝
总RNA
提取方法
Casuarina spp
twigs
total RNA
extraction method
作者简介
孙思，硕士，讲师，在职博士研究生．研究方向：分子植物病理．E-mail：sunsi@scau．edu．cn
通信作者：周而勋，博士，教授．研究方向：植物病原真菌及分子植物病理．E-mail：exzhou@SCHU．edu．cn
