植物学报 ›› 2001, Vol. 18 ›› Issue (06): 722-726.

• 研究论文 • 上一篇    下一篇

非洲菊花瓣总RNA提取方法的改进

张玉进 孟祥春 潘瑞炽 王小菁   

  1. (华南师范大学生物系,广东省植物发育生物工程重点实验室 广州 510631)
  • 收稿日期:2001-03-05 修回日期:2001-09-10 出版日期:2001-11-20 发布日期:2001-11-20
  • 通讯作者: 王小菁

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

ZHANG Yu-Jin MENG Xiang-Chun PAN Rui-Chi WANG Xiao-Jing   

  1. (Biology Department, South China Normal University, Guangdong Key Lab of Biotechnologyfor Plant Development, Guangzhou 510631)
  • Received:2001-03-05 Revised:2001-09-10 Online:2001-11-20 Published:2001-11-20
  • Contact: WANG Xiao-Jing

摘要: 利用异硫氰酸胍一步法和植物总RNA提取试剂盒(RNeasy Plant Mini Kit)提取非洲菊(Gerbera hybrida)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DEPC H2O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。

Abstract: 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 R NA 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.