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作 者 ：张玉进 孟祥春 潘瑞炽 王小菁

期 刊 ：植物学通报 2001年 18卷 06期 页码：722-726

关键词：非洲菊花瓣；异硫氰酸胍一步法；RNA提取试剂盒；总RNA；

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.

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