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为了获得高质量的枣RNA,本研究从所得RNA的纯度与浓度、操作时间和成本等方面比较了Trizol试剂法、传统CTAB法、热硼酸法、试剂盒法以及改良CTAB法5种方法提取枣不同组织总RNA的效果,并筛选出了最适合提取枣不同组织样品RNA的具体方法。结果表明:(1)在本研究试验的5种提取方法中,枣不同的组织RNA提取需要不同的方法。枣组培苗RNA提取的最佳方法是热硼酸法,枣芽和幼叶是试剂盒法,花和果实需要用改良CTAB法。(2)通过正交试验,得出枣树花和果实RNA提取适宜的改良CTAB法为在原有CTAB法的基础上水浴20 min、加入Na Ac的p H值为4.8、加入2倍体积的异丙醇。(3)枣RNA提取过程中的其他条件包括:RNA储存液保存的材料提取出的RNA质量最高,使用TBE缓冲液替代TAE进行电泳可以提高RNA的电泳亮度,4℃低温离心更有利于组培苗和芽的总RNA的提取,而叶片、花以及果实的RNA提取室温即可。
In order to obtain high quality jujube RNA, Trizol reagent method, traditional CTAB method, hot boric acid method, kit method and modified CTAB method were compared in terms of purity and concentration, operation time and cost of the obtained RNA Different tissues of total RNA, and screened out the most suitable method for extracting RNA from different tissue samples. The results showed that: (1) In the five extraction methods of this study, different methods of RNA extraction from different tissues of jujube were needed. Jujube tissue culture RNA extraction the best method is the hot boric acid method, jujube buds and young leaves is the kit method, flowers and fruits need to use improved CTAB method. (2) Through the orthogonal test, the RNA extracted from jujube tree and fruit was obtained by the modified CTAB method. In the original CTAB method, the water bath for 20 min, the pH value of Na Ac added to 4.8, Propanol. (3) Other conditions during the extraction of jujube RNA include: the RNA extracted from the RNA storage solution has the highest quality of RNA extracted. The use of TBE buffer instead of TAE for electrophoresis can improve the electrophoresis brightness of RNA. Low temperature centrifugation at 4 ° C is more conducive to tissue culture Mice and shoots are extracted from total RNA, while leaf, flower and fruit RNA extract at room temperature.