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顽拗植物龙眼果皮中总RNA的提取方法研究
作者姓名:王芳  董乐  林娈
作者单位:泉州师范学院化学与生命科学学院,泉州362000
基金项目:福建省科技厅青年科技人才创新项目“荒漠植物水通道蛋白基因分子克隆、定位、表达及功能研究”(2006F3113);福建省教育厅科技计划项目“龙眼多酚氧化酶eDNA的克隆及序列分析”(2007JA07151);泉州市科技局技术研究与开发项目“泉州市悬钩子属药用植物食用价值及其茶多酚含量的研究”(2007N6).
摘    要:【研究目的】为建立适于富含多糖和多酚等物质的龙眼果皮总RNA提取的方法;【方法】采用TRIzoL法、Tris-硼酸法和改良CTAB法提取龙眼果皮的总RNA。并通过凝胶电泳、紫外分光光度法检测提取的RNA样品的品质;【结果】改良CTAB法、Tris-硼酸法所提取的RNA经电泳检测。28SrRNA和18SrRNA条带清晰。RT-PCR得到的条带与目的片断大小一致。说明所提取的RNA纯度高、质量好;TRJZOL法所得到的RNA降解严重,没有得到完整的电泳条带;【结论】改良CTAB法和Tris-硼酸法适于富含多糖和多酚等物质的龙眼果皮总RNA的提取。

关 键 词:龙眼  果皮  总RNA  改良CTAB  Tris-硼酸法

The Method of RNA Isolation from Pericarp of Longan (Dimocarpus Longan Lour.) Fruits
Authors:Wang Fang  Dong Le  Lin Luan
Abstract:In order to establish a extraction method of total RNA from pericarp of longan Fruits in which polyphenols and polysaccharides are rich, the methods of TRIZOL , Tris - H3BO4 , and improved CTAB was used. The quality of total RNA was analyzed through gel eleetrophoresis and UV spectrometer. The resuits showed that two bands of 28S rRNA and 18S rRNA were appeared after gel eleetrophoresis with the method of improved CTAB and Tris - H3BO4 . The RNA samples were used to amplify eDNA through RT - PCR and the amplifications were observed in agarose gel by eleetrophoresis. The experimental results showed the RNA samples extracted with the method of TRIZOL obtained without gel eleetrophoresis band. The method of improved CTAB and Tris - H3BO4 are suited to extract the RNA of the periearp of longan Fruits.
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