非洲菊花瓣总RNA提取方法的改进

Modification of the Methods of Extracting Total RNA from Gerbera hybrida Corolla Tissue

利用异硫氰酸胍一步法和植物总RNA提取试剂盒 (RNeasyPlantMiniKit)提取非洲菊 (Gerberahy brida)花瓣中总RNA时存在多糖的干扰。实验中利用异硫氰酸胍一步法提取总RNA ,在RNA沉淀后 ,再次加入适量变性液 ,冻融 2次 ,再加热至 45℃使RNA完全溶解 ;试剂盒提取时 ,适当延长各提取步骤的离心时间 ,并增加DEPCH2 O溶解RNA的时间。通过以上操作方法的改进可排除多糖的干扰 ,得到高质量的总RNA。为进一步利用Northern杂交技术研究非洲菊中花色素苷基因的表达 ,进行花色改良提供了方便、有效的实验手段。

Using one_step Isothiocyanate method and RNeasy Plant Mini Kit, total RNA with high quality was extracted from Gerbera hybrida corolla tissue that is rich in polysaccharide. In order to remove polysaccharide interference in the procedure of RNA separation, we improved the method by adding denaturing_solution to dissolve the RNA precipitation and by prolonging the centrifugation time and dissolving time using DEPC H 2O. These provided a convenient and effective method for research on gene expression in anthocyanin biosynthesis using Northern hybridization and change in flower color. Finally, different methods for RNA separation aer compared.