摘要
[目的]设计一种简易安全的从农杆菌和酵母中提取RNA的方法。[方法]利用Triton X-100和SDS作为裂解液的主要成分,通过热激法裂解菌体,然后进行一系列的抽提、沉淀、洗涤,获得纯净的RNA;并用琼脂糖凝胶电泳及核酸检测仪检测总RNA的纯度、浓度及质量。[结果]该方法能高效裂解菌体,有效防止RNase污染,可以获得高质量的RNA。[结论]该研究成功的得到一种简易、安全提取农杆菌和酵母RNA的方法,为后续的基因表达分析奠定了技术基础。
[ Objective ] A simple and safe method for total RNA extraction from Agrobacterium and yeast was designed. [ Method ] Methods for RNA isolation is as following： bacterium was heated with lysis buffer（ Triton X-100 and SDS as the main composition）, then RNA was extrac- ted, precipitated and washed. The purity and quality of total RNA was identified by agarose gel electrophoresis and nucleic acid detection in- strument.. ~ Result] The method is efficient for pure RNA isolation without RNase contamination. [ Conclusion] Comparing to the previous methods, it was simple, high purity and safe and was worthy to be used for gene cloning and expression.
出处
《安徽农业科学》
CAS
2014年第7期1908-1909,1912,共3页
Journal of Anhui Agricultural Sciences
基金
国家自然科学基金项目(31370661)
关键词
农杆菌
酵母
RNA
提取方法
Agrobacterium tumefaciens
Saccharomyces cerevisiae
Total RNA
Extraction
